Fused heterocyclic compounds

ABSTRACT

Compounds, compositions and methods are provided that are useful in the treatment and/or prevention of a condition or disorder mediated by a G-protein coupled receptor. In particular, the compounds of the invention are useful in the treatment and/or prevention of eating disorders, obesity, anxiety disorders and mood disorders.

[0001] This application claims the benefit under 35 U.S.C. § 119 of U.S.provisional application No. 60/424,456, the contents of which are herebyincorporated by reference in their entirety.

FIELD OF THE INVENTION

[0002] The present invention relates to compounds, compositions andmethods useful in the treatment or prevention of conditions anddisorders associated with eating behavior, energy homeostasis andanxiety.

BACKGROUND OF THE INVENTION

[0003] G-protein coupled receptors play important roles in diversesignaling processes, including those involved with sensory and hormonalsignal transduction. Eating disorders, which represent a major healthconcern throughout the world, have been linked to GPCR regulation. Onthe one hand, disorders such as obesity, the excess deposition of fat inthe subcutaneous tissues, manifest themselves by an increase in bodyweight. Individuals who are obese often have, or are susceptible to,medical abnormalities including respiratory difficulties, cardiovasculardisease, diabetes and hypertension. On the other hand, disorders likecachexia, the general lack of nutrition and wasting associated withchronic disease and/or emotional disturbance, are associated with adecrease in body weight.

[0004] The neuropeptide melanin-concentrating hormone (MCH), a cyclichypothalamic peptide involved in the regulation of several functions inthe brain, has previously been found to be a major regulator of eatingbehavior and energy homeostasis. It has previously been determined thatMCH is the natural ligand for the 353-amino acid orphanG-protein-coupled-receptor (GPCR) termed SLC-1 (also known as GPR24).Subsequent to this determination, SLC-1, which is sequentiallyhomologous to the somatostatin receptors, is frequently referred to asmelanin-concentrating hormone receptor (MCH receptor, MCHR or MCHR1)(see Chambers et al., Nature 400:261-65 (1999); Saito et al., Nature400:265-69 (1999); and Saito et al., TEM 11(8):299-303 (2000)).

[0005] Compelling evidence exists that MCH is involved in regulation ofeating behavior. First, intracerebral administration of MCH in ratsresulted in stimulation of feeding. Next, mRNA corresponding to the MCHprecursor is up-regulated in the hypothalamus of genetically obese miceand of fasted animals. Finally, mice deficient in MCH are leaner andhave a decreased food intake relative to normal mice. MCH is believed toexert its activity by binding to MCHR, resulting in the mobilization ofintracellular calcium and a concomitant reduction in cAMP levels (seeChambers et al., Nature 400:261-65 (1999); Shimada et al. Nature396:670-74 (1998)). MCH also activates inwardly rectifying potassiumchannels, and MCHR has been found to interact with both Gαi protein andGαq protein (Saito et al., TEM 11(8):299-303 (2000)). Moreover, analysisof the tissue localization of MCHR indicates that it is expressed inthose regions of the brain involved in olfactory learning andreinforcement. The cumulative data suggest that modulators of MCHRshould have an effect on neuronal regulation of food intake (see Saitoet al., Nature 400:265-69 (1999)).

[0006] MCH has been shown to modulate behaviors other than feeding, suchas anxiety (Gonzales et al. (1996) Peptides 17:171-177; Monzon et al.(1999) Physiol. Behav. 67:813-817).

[0007] The identification of MCHR modulators is useful for the study ofphysiological processes mediated by MCHR and the development oftherapeutic agents for the treatment or prevention of conditions anddisorders associated with weight regulation, learning, anxiety and otherneuronal-related functions.

SUMMARY OF THE INVENTION

[0008] The present invention provides fused heterocyclic compounds andcompositions, and methods of use thereof to treat or prevent conditionsand disorders mediated by MCHR. In particular, the present inventionprovides compounds, compositions and methods for treating or preventingconditions and disorders associated with eating behavior, energyhomeostasis and anxiety.

[0009] The compounds provided herein have the formula (I):

[0010] wherein

[0011] represents a single or fused aryl or heteroaryl ring;

[0012] Q is —N(R)— or —N(R)—(C₁-C₃)alkylene-;

[0013] L¹ is a bond, (C₁-C₄)alkylene, (C₁-C₄)alkylenoxy and(C₁-C₄)alkylenamino;

[0014] L² is a bond, (C₁-C₄)alkylene, (C₂-C₄)alkenylene,(C₂-C₄)alkynylene, (C₁-C₄)alkylenoxy (e.g. —OCH₂CH₂—) or(C₁-C₄)alkylenamino (e.g. —NH—CH₂CH₂—);

[0015] R″ is hydrogen or (C₁-C₈)alkyl;

[0016] each R¹ is independently selected from the group consisting ofhalogen, (C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl,fluoro(C₁-C₄)alkyl, —OR⁵, —SR⁵, fluoro(C₁-C₄)alkoxy, aryl,aryl(C₁-C₄)alkyl, —NO₂, —NR⁵R⁶, —C(O)R⁵, —CO₂R⁵, —C(O)NR⁵R⁶,—N(R⁶)C(O)R⁵, —N(R⁶)CO₂R⁵, —N(R⁷)C(O)NR⁵R⁶, —S(O)_(m)NR⁵R⁶, —S(O)_(m)R⁵,—CN and —N(R⁶)S(O)_(m)R⁵;

[0017] R² and R³ are independently selected from the group consisting ofhydrogen, halogen, (C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl,fluoro(C₁-C₄)alkyl, —OR⁸, —SR⁸, fluoro(C₁-C₄)alkoxy, aryl,aryl(C₁-C₄)alkyl, —NO₂, —NR⁸R⁹, ═O, —C(O)R⁸, —CO₂R⁸, —C(O)NR⁸R⁹,—N(R⁹)C(O)R⁸, —N(R⁹)CO₂R⁸, —N(R¹⁰)C(O)NR⁸R⁹, —S(O)_(m)NR⁸R⁹,—S(O)_(m)R⁸, —CN and —N(R⁹)S(O)_(m)R⁸;

[0018] R⁴ is selected from the group consisting of hydrogen, —OR¹¹,—C(O)R¹¹, —CO₂R¹¹, —C(O)NR¹¹R¹², —CN, (C₁-C₄)alkyl and aryl;

[0019] X and Y are independently selected from the group consisting of(C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl, —CO₂R¹³ and —C(O)NR¹³R¹⁴;

[0020] optionally, X and Y may be combined to form a 3-, 4-, 5-, 6- or7-membered ring containing from 0 to 2 heteroatoms independentlyselected from the group consisting of N, O and S;

[0021] Z is selected from the group consisting of —OR¹⁵, —NR¹⁵R¹⁶,—NR¹⁵R¹⁸, —C(O)R¹⁵, —CO₂R¹⁵, —R¹⁸, —C(O)NR¹⁵R¹⁶, —C(O)NR¹⁵R¹⁸,—SO₂NR¹⁵R¹⁶, —SO₂NR¹⁵R¹⁸, —NR¹⁶SO₂R¹⁵, —N(R¹⁵)N(R¹⁶)SO₂R¹⁷,—C(O)N(R¹⁶)OR¹⁵, hydroxy(C₁-C₈)alkyl, fluoro(C₁-C₄)alkyl, heteroaryl,—C(═NOR¹⁵)NR¹⁶R¹⁷, —C(R¹⁶))═NOR¹⁵, —NR¹⁶(OR¹⁵), —C(O)NR¹⁷C(O)NR¹⁵R¹⁶,—NR¹⁷C(O)NR¹⁶C(O)R¹⁵ and —NR¹⁷C(O)NR¹⁵R¹⁶;

[0022] R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶ and R¹⁷ areindependently selected from the group consisting of hydrogen,(C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl, cyclo(C₃-C₆)alkyl,fluoro(C₁-C₄)alkyl, hetero(C₁-C₄)alkyl, cyclohetero(C₃-C₆)alkyl, aryland aryl(C₁-C₄)alkyl;

[0023] R¹⁸ is a 5- or 6-membered ring containing from 0 to 4 heteroatomsselected from the group consisting of N, O and S (e.g. tetrazole);

[0024] optionally, when two R groups selected from the group consistingof R⁵, R⁶, R⁸, R⁹, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶ and R¹⁷ are attached tothe same nitrogen atom, the R groups may be combined to form a 3-, 4-,5-, 6- or 7-membered ring containing the nitrogen atom and from 0 to 2additional heteroatoms selected from the group consisting of N, O and S;

[0025] the subscript m is 1 or 2; and

[0026] the subscript n is 0, 1 or 2.

[0027] In certain embodiments

[0028] represents a benzene, naphthalene, pyrrole, pyrazole, imidazole,pyrazine, oxazole, isoxazole, thiazole, furan, thiophene, pyridine,pyrimidine, benzothiazole, purine, benzimidazole, indole, isoquinoline,quinoxaline or quinoline ring.

[0029] In certain embodiments

[0030] represents a benzene ring.

[0031] In certain embodiments Q is —N(R)—.

[0032] In further embodiments, R³ is hydrogen or ═O.

[0033] In particular embodiments,

[0034] represents a benzene ring, R″ is hydrogen and R³ is hydrogen.

[0035] Further compounds provided herein have the formula (II):

[0036] wherein

[0037] L¹ is a bond, (C₁-C₄)alkylene, (C₁-C₄)alkylenoxy and(C₁-C₄)alkylenamino;

[0038] L² is a bond, (C₁-C₄)alkylene, (C₂-C₄)alkenylene,(C₂-C₄)alkynylene, (C₁-C₄)alkylenoxy (e.g —OCH₂CH₂—) or(C₁-C₄)alkylenamino (e.g. —NH—CH₂CH₂—);

[0039] R″ is hydrogen or (C₁-C₈)alkyl;

[0040] each R¹ is independently selected from the group consisting ofhalogen, (C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl,fluoro(C₁-C₄)alkyl, —OR⁵, —SR⁵, fluoro(C₁-C₄)alkoxy, aryl,aryl(C₁-C₄)alkyl, —NO₂, —NR⁵R⁶, —C(O)R⁵, —CO₂R⁵, —C(O)NR⁵R⁶,—N(R⁶)C(O)R⁵, —N(R⁶)CO₂R⁵, —N(R⁷)C(O)NR⁵R⁶, —S(O)_(m)NR⁵R⁶, —S(O)_(m)R⁵,—CN and —N(R⁶)S(O)_(m)R⁵;

[0041] R² is selected from the group consisting of hydrogen, halogen,(C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl, fluoro(C₁-C₄)alkyl, —OR⁸,—SR⁸, fluoro(C₁-C₄)alkoxy, aryl, aryl(C₁-C₄)alkyl, —NO₂, —NR⁸R⁹, ═O,—C(O)R⁸, —CO₂R⁸, —C(O)NR⁸R⁹, —N(R⁹)C(O)R⁸, —N(R⁹)CO₂R⁸,—N(R¹⁰)C(O)NR⁸R⁹, —S(O)_(m)NR⁸R⁹, —S(O)_(m)R⁸, —CN and —N(R⁹)S(O)_(m)R⁸;

[0042] R⁴ is selected from the group consisting of hydrogen, —OR¹¹,—C(O)R¹¹, —CO₂R¹¹, —C(O)NR¹¹R¹², —CN, (C₁-C₄)alkyl and aryl;

[0043] X and Y are independently selected from the group consisting of(C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl, —CO₂R¹³ and —C(O)NR¹³R¹⁴;

[0044] optionally, X and Y may be combined to form a 3-, 4-, 5-, 6- or7-membered ring containing from 0 to 2 heteroatoms independentlyselected from the group consisting of N, O and S;

[0045] Z is selected from the group consisting of —OR¹⁵, —NR¹⁵R¹⁶,—NR¹⁵R¹⁸, —C(O)R¹⁵, —CO₂R¹⁵, —R¹⁸, —C(O)NR¹⁵R¹⁶, —C(O)NR¹⁵R¹⁸,—SO₂NR¹⁵R¹⁶, —SO₂NR¹⁵R¹⁸ , NR¹⁶SO₂R¹⁵, —N(R¹⁵)N(R¹⁶)SO₂R¹⁷,—C(O)N(R¹⁶)OR¹⁵, hydroxy(C₁-C₈)alkyl, fluoro(C₁-C₄)alkyl, heteroaryl,—C(═NOR¹⁵)NR¹⁶R¹⁷, —C(R¹⁶)═NOR¹⁵, —NR¹⁶(OR¹⁵), —C(O)NR¹⁷C(O)NR¹⁵R¹⁶,—NR¹⁷C(O)NR¹⁶C(O)R¹⁵ and —NR¹⁷C(O)NR¹⁵R¹⁶;

[0046] R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹,R¹², R¹³, R¹⁴, R¹⁵, R¹⁶ and R¹⁷ areindependently selected from the group consisting of hydrogen,(C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl, cyclo(C₃-C₆)alkyl,fluoro(C₁-C₄)alkyl, hetero(C₁-C₄)alkyl, cyclohetero(C₃-C₆)alkyl, aryland aryl(C₁-C₄)alkyl;

[0047] R¹⁸ is a 5- or 6-membered ring containing from 0 to 4 heteroatomsselected from the group consisting of N, O and S (e.g. tetrazole);

[0048] optionally, when two R groups selected from the group consistingof R⁵, R⁶, R⁸, R⁹, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶ and R¹⁷ are attached tothe same nitrogen atom, the R groups may be combined to form a 3-, 4-,5-, 6- or 7-membered ring containing the nitrogen atom and from 0 to 2additional heteroatoms selected from the group consisting of N, O and S;

[0049] the subscript m is 1 or 2; and

[0050] the subscript n is 0, 1 or 2.

[0051] The compounds provided in the above formulas are meant to includeall pharmaceutically acceptable salts, hydrates, solvates or prodrugsthereof.

[0052] The pharmaceutical compositions provided herein comprise apharmaceutically acceptable carrier or excipient in combination with acompound of formula I or II.

[0053] Methods for treating or preventing a condition or disorderselected from the group consisting of obesity, an eating disorder, ananxiety disorder and a mood disorder are provided herein. The methodscomprise administering to a subject in need thereof a therapeuticallyeffective amount of one of the foregoing compounds or pharmaceuticalcompositions.

[0054] Other objects, features and advantages of the present inventionwill become apparent to those skilled in the art from the followingdescription and claims.

BRIEF DESCRIPTION OF THE DRAWINGS

[0055]FIG. 1 provides the structures of exemplary compounds of theinvention.

DETAILED DESCRIPTION OF THE INVENTION

[0056] Abbreviations and Definitions

[0057] The abbreviations used herein are conventional, unless otherwisedefined.

[0058] The term “MCHR” refers to the melanin-concentrating hormonereceptor protein 1 (MCHR1), unless otherwise stated.

[0059] The terms “treat”, “treating” and “treatment” refer to a methodof alleviating or abrogating a disease and/or its attendant symptoms.

[0060] The terms “prevent”, “preventing” and “prevention” refer to amethod of decreasing the probability or eliminating the possibility thata disease will be contracted.

[0061] As used herein, the term “MCHR-mediated condition or disorder”and the like refers to a condition or disorder characterized byinappropriate, e.g., less than or greater than normal, MCHR activity. AnMCHR-mediated condition or disorder may be completely or partiallymediated by inappropriate MCHR activity. However, an MCHR-mediatedcondition or disorder is one in which modulation of MCHR results in someeffect on the underlying condition or disease (e.g., an MCHR antagonistresults in some improvement in patient well-being in at least somepatients). Exemplary MCHR-mediated conditions and disorders includeobesity, eating disorders and other behavioral disorders, such asanxiety disorders and mood disorders.

[0062] The term “therapeutically effective amount” refers to that amountof the compound being administered sufficient to prevent development ofor alleviate to some extent one or more of the symptoms of the conditionor disorder being treated.

[0063] As used herein, the term “obesity” refers to the excessiveaccumulation of body fat. Obesity may have genetic, environmental (e.g.,expending less energy than is consumed) and regulatory determinants.Cardiovascular disorders, lipid disorders and metabolic disorders, suchas hypertension, hyperlidemia, coronary artery disease and diabetes, arecommonly associated with obesity.

[0064] As used herein, the terms “eating disorder”, “feeding disorder”,and the like refer to an emotional and/or behavioral disturbanceassociated with an excessive decrease in body weight and/orinappropriate efforts to avoid weight gain, e.g., fasting, self-inducedvomiting, laxative or diuretic abuse. Depression is commonly associatedwith eating disorders. Exemplary eating disorders include anorexianervosa and bulimia.

[0065] As used herein, the term “anxiety disorder” refers to anemotional and/or behavioral disturbance characterized by persistent andpervasive worry or restlessness, tension or irritability about, e.g.,health, work, money or family, for no clear reason. An anxiety disordermay be accompanied by tachycardia or dyspnea. Exemplary anxietydisorders include anxiety, generalized anxiety disorder, panic attacks,panic disorder and obsessive-compulsive disorder (OCD).

[0066] As used herein, the term “mood disorder” refers to an emotionaland/or behavioral disturbance characterized by persistent and pervasivebouts of euphoria and/or depression. Exemplary mood disorders includedepression and bipolar disorders. Anxiety is frequently associated withmood disorders, such as depression.

[0067] The term “modulate” refers to the ability of a compound toincrease or decrease the function, or activity, of MCHR. Modulation, asdescribed herein, includes the antagonism or agonism of MCHR, eitherdirectly or indirectly. Antagonist are compounds that, e.g., partiallyor totally block stimulation, decrease, prevent, delay activation,inactivate, inhibit, desensitize, or down-regulate signal transduction.Agonists are compounds that, e.g., stimulate, increase, activate, open,facilitate, enhance activation, sensitize or up-regulate signaltransduction.

[0068] The term “alkyl,” by itself or as part of another substituent,means, unless otherwise stated, a straight or branched chain, or cyclichydrocarbon radical, or combination thereof, which is fully saturated,having the number of carbon atoms designated (i.e. C₁-C₈ means one toeight carbons). Examples of alkyl groups include methyl, ethyl,n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, cyclohexyl,(cyclohexyl)methyl, cyclopropylmethyl, homologs and isomers of, forexample, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. As usedherein, (C₁-C₈)alkyl refers to an alkyl group having from one to eightcarbon atoms and includes, e.g., (C₁-C₄)alkyl.

[0069] The term “alkenyl”, by itself or as part of another substituent,means a straight or branched chain, or cyclic hydrocarbon radical, orcombination thereof, which may be mono- or polyunsaturated, having thenumber of carbon atoms designated (i.e. C₂-C₈ means two to eightcarbons) and one or more double bonds. Examples of alkenyl groupsinclude vinyl, allyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl),2,4-pentadienyl, 3-(1,4-pentadienyl) and higher homologs and isomersthereof.

[0070] The term “alkynyl”, by itself or as part of another substituent,means a straight or branched chain hydrocarbon radical, or combinationthereof, which may be mono- or polyunsaturated, having the number ofcarbon atoms designated (i.e. C₂-C₈ means two to eight carbons) and oneor more triple bonds. Examples of alkynyl groups include ethynyl, 1- and3-propynyl, 3-butynyl and higher homologs and isomers thereof.

[0071] The term “alkylene” by itself or as part of another substituentmeans a divalent radical derived from alkyl, as exemplified by—CH₂CH₂CH₂CH₂—. Typically, an alkyl (or alkylene) group will have from 1to 24 carbon atoms, with those groups having 10 or fewer carbon atomsbeing preferred in the present invention. A “lower alkyl” or “loweralkylene” is a shorter chain alkyl or alkylene group, generally havingseven or fewer carbon atoms.

[0072] The terms “alkoxy,” “alkylamino” and “alkylthio” (or thioalkoxy)are used in their conventional sense, and refer to those alkyl groupsattached to the remainder of the molecule via an oxygen atom, an aminogroup, or a sulfur atom, respectively.

[0073] The term “heteroalkyl,” by itself or in combination with anotherterm, means, unless otherwise stated, a stable straight or branchedchain, or cyclic hydrocarbon radical, or combinations thereof,consisting of the stated number of carbon atoms and from one to threeheteroatoms selected from the group consisting of O, N, Si and S,wherein the nitrogen and sulfur atoms may optionally be oxidized and thenitrogen heteroatom may optionally be quaternized. The heteroatom(s) O,N and S may be placed at any interior position of the heteroalkyl group.The heteroatom Si may be placed at any position of the heteroalkylgroup, including the position at which the alkyl group is attached tothe remainder of the molecule. Examples include —CH₂—CH₂—O—CH₃,—CH₂—CH₂—NH—CH₃, —CH₂—CH₂—N(CH₃)—CH₃, —CH₂—S—CH₂—CH₃, —CH₂—CH₂,—S(O)—CH₃, —CH₂—CH₂—S(O)₂—CH₃, —CH═CH—O—CH₃, —Si(CH₃)₃, —CH₂—CH═N—OCH₃,and —CH═CH—N(CH₃)—CH₃. Up to two heteroatoms may be consecutive, suchas, for example, —CH₂—NH—OCH₃ and —CH₂—O—Si(CH₃)₃

[0074] Similarly, the term “heteroalkylene” by itself or as part ofanother substituent means a divalent radical derived from heteroalkyl,as exemplified by —CH₂—CH₂—S—CH₂CH₂— and —CH₂—S—CH₂—CH₂—NH—CH₂—. Forheteroalkylene groups, heteroatoms can also occupy either or both of thechain termini (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino,alkylenediamino, and the like). Still further, for alkylene andheteroalkylene linking groups, no orientation of the linking group isimplied.

[0075] The terms “cycloalkyl” and “heterocycloalkyl”, by themselves orin combination with other terms, represent, unless otherwise stated,cyclic versions of “alkyl” and “heteroalkyl”, respectively. Accordingly,a cycloalkyl group has the number of carbon atoms designated (i.e.,C₃-C₈ means three to eight carbons) and a heterocycloalkyl groupconsists of the number of atoms designated (i.e., C₂-C₈ means two toeight carbons) and from one to three heteroatoms selected from the groupconsisting of O, N, Si and S, and wherein the nitrogen and sulfur atomsmay optionally be oxidized and the nitrogen heteroatom may optionally bequaternized. Additionally, for heterocycloalkyl, a heteroatom can occupythe position at which the heterocycle is attached to the remainder ofthe molecule. Examples of cycloalkyl include cyclopentyl, cyclohexyl,1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Examples ofheterocycloalkyl include 1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl,2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl,tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothien-2-yl,tetrahydrothien-3-yl, 1-piperazinyl, 2-piperazinyl, and the like.

[0076] The terms “halo” and “halogen,” by themselves or as part ofanother substituent, mean, unless otherwise stated, a fluorine,chlorine, bromine, or iodine atom. Additionally, terms such as“haloalkyl,” are meant to include alkyl substituted with halogen atoms,which can be the same or different, in a number ranging from one to(2m′+1), where m′ is the total number of carbon atoms in the alkylgroup. For example, the term “halo(C₁-C₄)alkyl” is mean to includetrifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, andthe like. Thus, the term “haloalkyl” includes monohaloalkyl (alkylsubstituted with one halogen atom) and polyhaloalkyl (alkyl substitutedwith halogen atoms in a number ranging from two to (2m′+1) halogenatoms, where m′ is the total number of carbon atoms in the alkyl group).Accordingly, the term “fluoro(C₁-C₄)alkyl” includes fluoromethyl,trifluoromethyl, 2,2,2-trifluoroethyl, pentafluoroethyl,1,1-difluoroethyl, and the like. The term “perhaloalkyl” means, unlessotherwise stated, alkyl substituted with (2m′+1) halogen atoms, where m′is the total number of carbon atoms in the alkyl group. For example theterm “perhalo(C₁-C₄)alkyl” is meant to include trifluoromethyl,pentachloroethyl, 1,1,1-trifluoro-2-bromo-2-chloroethyl, and the like.

[0077] The term “aryl” means, unless otherwise stated, apolyunsaturated, typically aromatic, hydrocarbon substituent which canbe a single ring or multiple rings (up to three rings) which are fusedtogether or linked covalently. Non-limiting examples of aryl groupsinclude phenyl, 1-naphthyl, 2-naphthyl and 4-biphenyl.

[0078] The term “heteroaryl” refers to aryl groups (or rings) thatcontain from zero to four heteroatoms selected from N, O, and S, whereinthe nitrogen and sulfur atoms are optionally oxidized and the nitrogenheteroatom are optionally quaternized. A heteroaryl group can beattached to the remainder of the molecule through a heteroatom.Non-limiting examples of heteroaryl groups include 1-pyrrolyl,2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl,pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl,3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl,5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl,3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl,purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl,2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl and 6-quinolyl.

[0079] For brevity, the term “aryl” when used in combination with otherterms (e.g., aryloxy, arylthioxy, arylalkyl) includes both aryl andheteroaryl rings as defined above. Thus, the term “arylalkyl” is meantto include those radicals in which an aryl group is attached to an alkylgroup (e.g., benzyl, phenethyl, pyridylmethyl, and the like) includingthose alkyl groups in which a carbon atom (e.g., a methylene group) hasbeen replaced by, for example, an oxygen atom (e.g., phenoxymethyl,2-pyridyloxymethyl, 3-(1-naphthyloxy)propyl, and the like).

[0080] Each of the above terms (e.g., “alkyl,” “heteroalkyl,” “aryl” and“heteroaryl”) is meant to include both substituted and unsubstitutedforms of the indicated radical. Preferred substituents for each type ofradical are provided below.

[0081] Substituents for the alkyl and heteroalkyl radicals (as well asthose groups referred to as alkylene, alkenyl, heteroalkylene,heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl andheterocycloalkenyl) can be a variety of groups selected from: —OR′, ═O,═NR′, ═N—OR′, —NR′R″, —SR′, halogen, —SiR′R″R′″, —OC(O)R′, —C(O)R′,—CO₂R′, —CONR′R″, —OC(O)NR′R″, —NR″C(O)R′, —NR′—C(O)NR″R′″,—NR′—SO₂NR″R′″, —NR″CO₂R′, —NH—C(NH₂)═NH, —NR′C(NH₂)═NH, —NH—C(NH₂)═NR′,—S(O)R′, —SO₂R′, —SO₂NR′R″, —NR″SO₂R′, —CN and —NO₂, in a number rangingfrom zero to three, with those groups having zero, one or twosubstituents being particularly preferred. R′, R″ and R′″ eachindependently refer to hydrogen, unsubstituted (C₁-C₈)alkyl andheteroalkyl, fluoro(C₁-C₄)alkyl, unsubstituted aryl, aryl substitutedwith one to three halogens, unsubstituted alkyl, alkoxy or thioalkoxygroups, or aryl(C₁-C₄)alkyl groups. When R′ and R″ are attached to thesame nitrogen atom, they can be combined with the nitrogen atom to forma 5-, 6- or 7-membered ring. For example, —NR′R″ is meant to include1-pyrrolidinyl and 4-morpholinyl. Typically, an alkyl or heteroalkylgroup will have from zero to three substituents, with those groupshaving two or fewer substituents being preferred in the presentinvention. More preferably, an alkyl or heteroalkyl radical will beunsubstituted or monosubstituted. Most preferably, an alkyl orheteroalkyl radical will be unsubstituted. From the above discussion ofsubstituents, one of skill in the art will understand that the term“alkyl” is meant to include groups such as trihaloalkyl (e.g., —CF₃ and—CH₂CF₃).

[0082] Preferred substituents for the alkyl and heteroalkyl radicals areselected from: —OR′, ═O, —NR′R″, —SR′, halogen, —SiR′R″R′″, —OC(O)R′,—C(O)R′, —CO₂R′, —CONR′R″, —OC(O)NR′R″, —NR″C(O)R′, —NR″CO₂R′,—NR′—SO₂NR″R′″, —S(O)R′, —SO₂R′, —SO₂NR′R″, —NR″SO₂R′, —CN and —NO₂,where R′ and R″ are as defined above. Further preferred substituents areselected from: —OR′, ═O, —NR′R″, halogen, —OC(O)R′, —CO₂R′, —CONR′R″,—OC(O)NR′R″, —NR″C(O)R′, —NR″CO₂R′, —NR′—SO₂NR″R′″, —SO₂R′, —SO₂NR′R″,—NR″SO₂R′, —CN and —NO₂

[0083] Similarly, substituents for the aryl and heteroaryl groups arevaried and selected from: halogen, —OR′, —OC(O)R′, —NR′R″, —SR′, —R′,—CN, —NO₂, —CO₂R′, —CONR′R″, —C(O)R′, —OC(O)NR′R″, —NR″C(O)R′,—NR″CO₂R′, —NR′—C(O)NR″R′″, —NR′—SO₂NR″R′″, —NH—C(NH₂)═NH,—NR′C(NH₂)═NH, —NH—C(NH₂)═NR′, —S(O)R′, —SO₂R′, —SO₂NR′R″, —NR″SO₂R′,—N₃, —CH(Ph)₂, perfluoro(C₁-C₄)alkoxy and perfluoro(C₁-C₄)alkyl, in anumber ranging from zero to the total number of open valences on thearomatic ring system; and where R′, R″ and R′″ are independentlyselected from hydrogen, (C₁-C₈)alkyl and heteroalkyl, unsubstituted aryland heteroaryl, (unsubstituted aryl)-(C₁-C₄)alkyl and (unsubstitutedaryl)oxy-(C₁-C₄)alkyl. Typically, an aryl or heteroaryl group will havefrom zero to three substituents, with those groups having two or fewersubstituents being preferred in the present invention. More preferably,an aryl or heteroaryl group will be unsubstituted or monosubstituted.Most preferably, an aryl or heteroaryl group will be unsubstituted.

[0084] Preferred substituents for aryl and heteroaryl groups areselected from: halogen, —OR′, —OC(O)R′, —NR′R″, —SR′, —R′, —CN, —NO₂,—CO₂R′, —CONR′R″, —C(O)R′, —OC(O)NR′R″, —NR″C(O)R′, —S(O)R′, —SO₂R′,—SO₂NR′R″, —NR″SO₂R′, —N₃, —CH(Ph)₂, perfluoro(C₁-C₄)alkoxy andperfluoro(C₁-C₄)alkyl, where R′ and R″ are as defined above. Furtherpreferred substituents are selected from: halogen, —OR′, —OC(O)R′,—NR′R″, —R′, —CN, —NO₂, —CO₂R′, —CONR′R″, —NR″C(O)R′, —SO₂R′, —SO₂NR′R″,—NR″SO₂R′, perfluoro(C₁-C₄)alkoxy and perfluoro(C₁-C₄)alkyl

[0085] As used herein, the substituent —CO₂H, includes bioisostericreplacements therefor, such as:

[0086] and the like. See, e.g., The Practice of Medicinal Chemistry;Wermuth, C. G., Ed.; Academic Press: New York, 1996; p. 203.

[0087] Two of the substituents on adjacent atoms of the aryl ring mayoptionally be replaced with a substituent of the formula-T-C(O)—(CH₂)_(q)—U—, wherein T and U are independently —NH—, —O—, —CH₂—or a single bond, and q is an integer of from 0 to 2. Alternatively, twoof the substituents on adjacent atoms of the aryl ring may optionally bereplaced with a substituent of the formula -A-(CH₂)_(r)—B—, wherein Aand B are independently —CH₂—, —O—, —NH—, —S—, —S(O)—, —SO₂—, —SO₂NR′—or a single bond, and r is an integer of from 1 to 3. One of the singlebonds of the new ring so formed may optionally be replaced with a doublebond. Alternatively, two of the substituents on adjacent atoms of thearyl ring may optionally be replaced with a substituent of the formula—(CH₂)_(s)—X—(CH₂)_(t)—, where s and t are independently integers offrom 0 to 3, and X is —O—, —NR′—, —S—, —S(O)—, —SO₂—, or —SO₂NR′—. Thesubstituent R′ in —NR′— and —SO₂NR′— is selected from the groupconsisting of hydrogen or unsubstituted (C₁-C₆)alkyl.

[0088] As used herein, the term “heteroatom” is meant to include oxygen(O), nitrogen (N), sulfur (S) and silicon (Si).

[0089] The term “pharmaceutically acceptable salts” is meant to includesalts of the active compounds which are prepared with relativelynontoxic acids or bases, depending on the particular substituents foundon the compounds described herein. When compounds of the presentinvention contain relatively acidic functionalities, base addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired base, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable base additionsalts include sodium, potassium, calcium, ammonium, organic amino, ormagnesium salt, or a similar salt. When compounds of the presentinvention contain relatively basic functionalities, acid addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, oxalic, maleic, malonic, benzoic,succinic, suberic, fumaric, mandelic, phthalic, benzenesulfonic,p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Alsoincluded are salts of amino acids such as arginate and the like, andsalts of organic acids like glucuronic or galactunoric acids and thelike (see, for example, Berge, S. M., et al. (1977) J. Pharm. Sci.66:1-19). Certain specific compounds of the present invention containboth basic and acidic functionalities that allow the compounds to beconverted into either base or acid addition salts.

[0090] The neutral forms of the compounds may be regenerated bycontacting the salt with a base or acid and isolating the parentcompound in the conventional manner. The parent form of the compounddiffers from the various salt forms in certain physical properties, suchas solubility in polar solvents, but otherwise the salts are equivalentto the parent form of the compound for the purposes of the presentinvention.

[0091] In addition to salt forms, the present invention providescompounds which are in a prodrug form. Prodrugs of the compoundsdescribed herein are those compounds that readily undergo chemicalchanges under physiological conditions to provide the compounds of thepresent invention. Additionally, prodrugs can be converted to thecompounds of the present invention by chemical or biochemical methods inan ex vivo environment. For example, prodrugs can be slowly converted tothe compounds of the present invention when placed in a transdermalpatch reservoir with a suitable enzyme or chemical reagent. Prodrugs areoften useful because, in some situations, they may be easier toadminister than the parent drug. They may, for instance, be bioavailableby oral administration whereas the parent drug is not. The prodrug mayalso have improved solubility in pharmacological compositions over theparent drug. A wide variety of prodrug derivatives are known in the art,such as those that rely on hydrolytic cleavage or oxidative activationof the prodrug. An example, without limitation, of a prodrug would be acompound of the present invention which is administered as an ester (the“prodrug”), but then is metabolically hydrolyzed to the carboxylic acid,the active entity. Additional examples include peptidyl derivatives of acompound of the invention.

[0092] Certain compounds of the present invention can exist inunsolvated forms as well as solvated forms, including hydrated forms. Ingeneral, the solvated forms are equivalent to unsolvated forms and areintended to be encompassed within the scope of the present invention.Certain compounds of the present invention may exist in multiplecrystalline or amorphous forms. In general, all physical forms areequivalent for the uses contemplated by the present invention and areintended to be within the scope of the present invention.

[0093] Certain compounds of the present invention possess asymmetriccarbon atoms (optical centers) or double bonds; the racemates,enantiomers, diastereomers, geometric isomers and individual isomers areall intended to be encompassed within the scope of the presentinvention.

[0094] The compounds of the present invention may also contain unnaturalproportions of atomic isotopes at one or more of the atoms thatconstitute such compounds. For example, the compounds may beradiolabeled with radioactive isotopes, such as for example tritium(³H), iodine-125 (¹²⁵I) or carbon-14 (¹⁴C). All isotopic variations ofthe compounds of the present invention, whether radioactive or not, areintended to be encompassed within the scope of the present invention.

EMBODIMENTS OF THE INVENTION

[0095] MCHR (GenBank Accession No. U71092) is expressed in brain, atmoderate levels in the eye and skeletal muscle, and in low levels intongue and the pituitary gland. Evidence suggests that MCHR is involvedin, inter alia, olfactory learning, regulation of feeding behavior andenergy metabolism, regulation of thehypothalmic-pituitary-adrenocortical axis following stress, arousal andthe sensation of anxiety (Saito et al., TEM 11(8):299-303 (2000)). Thecompounds of the present invention inhibit MCHR activity, and thus, areuseful in, for example, the treatment or prevention of disordersassociated with these processes.

[0096] Compounds

[0097] In one aspect, the present invention provides compoundsrepresented by the formula (I):

[0098] wherein

[0099] or a pharmaceutically acceptable salt, hydrate, solvate orprodrug thereof. In formula I,

[0100] represents a single or fused aryl or heteroaryl ring. Forinstance,

[0101] can represent benzene, naphthalene, pyrrole, pyrazole, imidazole,pyrazine, oxazole, isoxazole, thiazole, furan, thiophene, pyridine,pyrimidine, benzothiazole, purine, benzimidazole, indole, isoquinoline,quinoxaline or quinoline ring. In preferred embodiments,

[0102] represents benzene.

[0103] The symbol Q represents —N(R)— or —N(R)—(C₁-C₃)alkylene-. Incertain embodiments the symbol Q represents —N(R)—.

[0104] The symbol R represents

[0105] The symbol L¹ represents a divalent linkage selected from a bond,(C₁-C₄)alkylene, (C₁-C₄)alkylenoxy and (C₁-C₄)alkylenamino. Exemplary L¹groups are a single bond, methylene, ethylene, n-propylene andn-butylene. The symbol L² represents a divelent linkage selected from abond, (C₁-C₄)alkylene, (C₂-C₄)alkenylene, (C₂-C₄)alkynylene,(C₁-C₄)alkylenoxy and (C₁-C₄)alkylenamino. Exemplary L² groups are a asingle bond, methylene, ethylene, n-propylene and n-butylene.

[0106] The letters X and Y represent independently (C₁-C₈)alkyl,(C₂-C₈)alkenyl, (C₂-C₈)alkynyl, —CO₂R¹³ or —C(O)NR¹³R¹⁴. Optionally, Xand Y may be combined to form a 3-, 4-, 5-, 6- or 7-membered ringcontaining from 0 to 2 heteroatoms selected from N, O and S.

[0107] The letter Z represents —OR¹², —N¹²R¹³, —CO₂R¹², —R¹⁵,—C(O)NR¹²R¹³, —C(O)NR¹²R¹⁵, —SO₂NR¹²R¹³, —NR¹³SO₂R¹⁵,—N(R¹²)N(R¹³)SO₂R¹⁴, —C(O)N(R¹³)OR¹², fluoro(C₁-C₄)alkyl, heteroaryl,—C(═NOR¹²)NR¹³R¹⁴, —C(R¹³)═NOR¹², —NR¹³(OR¹²), —C(O)NR¹⁴C(O)NR¹²R¹³,—NR¹⁴C(O)NR¹³C(O)R¹² and —NR¹⁴C(O)NR¹²R¹³. Exemplary —C(X)(Y)(L²Z)groups are:

[0108] R″ is hydrogen or (C₁-C₈)alkyl.

[0109] Each R¹ is independently halogen, (C₁-C₈)alkyl, (C₂-C₈)alkenyl,(C₂-C₈)alkynyl, fluoro(C₁-C₄)alkyl, —OR⁵, —SR⁵, fluoro(C₁-C₄)alkoxy,aryl, aryl(C₁-C₄)alkyl, —NO₂, —NR⁵R⁶, —C(O)R⁵, —CO₂R⁵, —C(O)NR⁵R⁶,—N(R⁶)C(O)R⁵, —N(R⁶)CO₂R⁵, —N(R⁷)C(O)NR⁵R⁶, —S(O)_(m)NR⁵R⁶, —S(O)_(m)R⁵,—CN or —N(R⁶)S(O)_(m)R⁵. Exemplary R¹ groups are Cl and CF₃.

[0110] R² and R³ are independently selected from hydrogen, halogen,(C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl, fluoro(C₁-C₄)alkyl, —OR⁸,—SR⁸, fluor(C₁-C₄)alkoxy, aryl, aryl(C₁-C₄)alkyl, —NO₂, —NR⁸R⁹, ═O,—C(O)R⁸, —CO₂R⁸, —C(O)NR⁸R⁹, —N(R⁹)C(O)R⁸, —N(R⁹)CO₂R⁸,—N(R¹⁰)C(O)NR⁸R⁹, —S(O)_(m)NR⁸R⁹, —S(O)_(m)R⁸, —CN and —N(R⁹)S(O)R⁸.Exemplary R² groups are methyl, isopropyl, trifluoromethyl, hydroxy,methoxy, hydroxymethyl, trifluoromethoxy, phenyl and ═O. In certainembodiments, R³ is hydrogen or ═O.

[0111] R⁴ is hydrogen, —OR¹¹, —C(O)R¹¹, —CO₂R¹¹, —C(O)NR¹¹R¹², —CN,(C₁-C₄)alkyl or aryl.

[0112] R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁶ and R¹⁷ areindependently selected from hydrogen, (C₁-C₈)alkyl, (C₂-C₈)alkenyl,(C₂-C₈)alkynyl, fluoro(C₁-C₄)alkyl, hetero(C₁-C₄)alkyl, aryl andaryl(C₁-C₄)alkyl and R¹⁸ is a 5- or 6-membered ring containing from 1 to3 heteroatoms selected from N, O and S. The subscript m is 1 or 2 andthe subscript n is 0, 1 or 2. Optionally, when two R groups selectedfrom the group consisting of R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³,R¹⁴, R¹⁵, R¹⁶, R¹⁷ and R¹⁸ are attached to the same nitrogen atom, the Rgroups may be combined to form a 3-, 4-, 5-, 6- or 7-membered ringcontaining the nitrogen atom and from 0 to 2 additional heteroatomsselected from N, O and S.

[0113] In particular embodiments,

[0114] represents a benzene ring, R″ is hydrogen and R³ is hydrogen.

[0115] In another aspect, the present invention provides compounds offormula (II):

[0116] or a pharmaceutically acceptable salt, hydrate, solvate orprodrug thereof. In formula II, the symbol L¹ represents a divalentlinkage selected from a bond, (C₁-C₄)alkylene, (C₁-C₄)alkylenoxy and(C₁-C₄)alkylenamino. Exemplary L¹ groups are a single bond, methylene,ethylene, n-propylene and n-butylene. The symbol L² represents adivalent linkage selected from a bond, (C₁-C₄)alkylene,(C₂-C₄)alkenylene, (C₂-C₄)alkynylene, (C₁-C₄)alkylenoxy and(C₁-C₄)alkylenamino. Exemplary L² groups are a single bond, methylene,ethylene, n-propylene and n-butylene.

[0117] The letters X and Y represent independently (C₁-C₈)alkyl,(C₂-C₈)alkenyl, (C₂-C₈)alkynyl, —CO₂R¹³ or —C(O)NR¹³R¹⁴. Optionally, Xand Y may be combined to form a 3-, 4-, 5-, 6- or 7-membered ringcontaining from 0 to 2 heteroatoms selected from N, O and S.

[0118] The letter Z represents —OR¹², —NR¹²R¹³, —CO₂R¹³, —R¹⁵,—C(O)NR¹²R¹³, —C(O)NR¹²R¹⁵, —SO₂NR¹³R¹³, —NR¹³SO₂R¹⁵,—N(R¹²)N(R¹³)SO₂R¹⁴, —C(O)N(R¹³)OR¹², fluoro(C₁-C₄)alkyl, heteroaryl,—C(═NOR¹²)NR¹³R¹⁴, —C(R³ )═NOR¹², —NR¹³(OR¹²), —C(O)NR¹⁴C(O)NR¹²R¹³,—NR¹⁴C(O)NR¹³C(O)R¹² and —NR¹⁴C(O)NR¹²R¹³. Exemplary —C(X)(Y)(L²Z)groups are:

[0119] R″ is hydrogen or (C₁-C₈)alkyl.

[0120] Each R¹ is independently halogen, (C₁-C₈)alkyl, (C₂-C₈)alkenyl,(C₂-C₈)alkynyl, fluoro(C₁-C₄)alkyl, —OR⁵, —SR⁵, fluoro(C₁-C₄)alkoxy,aryl, aryl(C₁-C₄)atkyl, —NO₂, —NR⁵R⁶, —C(O)R⁵, —CO₂R⁵, —C(O)NR⁵R⁶,—N(R⁶)C(O)R⁵, —N(R⁷)C(O)NR⁵R⁶, —S(O)_(m)NR⁵R⁶, —S(O)_(m)R⁵, —CN or—N(R⁶)S(O)_(m)R⁵. Exemplary R¹ groups are Cl and CF₃.

[0121] R² is selected from hydrogen, halogen, (C₁-C₈)alkyl,(C₂-C₈)alkenyl, (C₂-C₈)alkynyl, fluoro(C₁-C₄)alkyl, —OR⁸, —SR⁸,fluoro(C₁-C₄)alkoxy, aryl, aryl(C₁-C₄)alkyl, —NO₂, —NR⁸R⁹, ═O, —C(O)R⁸,—CO₂R⁸, —C(O)NR⁸R⁹, —N(R⁹)C(O)R⁸, —N(R⁹)CO₂R⁸, —N(R¹⁰)C(O)NR⁸R⁹,—S(O)_(m)NR⁸R⁹, —S(O)_(m)R⁸, —CN and —N(R⁹)S(O)_(m)R⁸. Exemplary R²groups are methyl, isopropyl, trifluoromethyl, hydroxy, methoxy,hydroxymethyl, trifluoromethoxy, phenyl and ═O.

[0122] R⁴ is hydrogen, —OR¹¹, —C(O)R¹¹, —CO₂R¹¹, —C(O)NR¹¹R¹², —CN,(C₁-C₄)alkyl or aryl.

[0123] R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁶ and R¹⁷ areindependently selected from hydrogen, (C₁-C₈)alkyl, (C₂-C₈)alkenyl,(C₂-C₈)alkynyl, fluoro(C₁-C₄)alkyl, hetero(C₁-C₄)alkyl, aryl andaryl(C₁-C₄)alkyl and R¹⁸ is a 5- or 6-membered ring containing from 1 to3 heteroatoms selected from N, O and S. The subscript m is 1 or 2 andthe subscript n is 0, 1 or 2. Optionally, when two R groups selectedfrom the group consisting R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵,R¹⁶, R¹⁷ and R¹⁸ are attached to the same nitrogen atom, the R groupsmay be combined to form a 3-, 4-, 5-, 6- or 7-membered ring containingthe nitrogen atom and from 0 to 2 additional heteroatoms selected fromN, O and S.

[0124] Compounds of the invention feature apyrido[4,3-b]carbazole-derived ring, minimally substituted at the 2- and11-positions. The ring numbering system used herein is illustratedbelow.

[0125] One of skill in the art will understand that formula IIencompasses two enantiomers. The enantiomers have the structuralorientations represented by the following formulae:

[0126] Within formula I or II above, a number of groups of embodimentsare preferred, described below.

[0127] In one group of preferred embodiments, L¹ is (C₁-C₄)alkylene. Ina preferred embodiment, L¹ is unsubstituted (C₁-C₄)alkylene or—(CH₂)_(p)—, wherein the subscript p is an integer of from 1 to 4. In afurther preferred embodiment, p is 1, 2 or 3. In a still furtherpreferred embodiment, p is 2 or 3. In a particularly preferredembodiment, p is 2.

[0128] One group of preferred embodiments is represented by the formula(III):

[0129] In a preferred embodiment, X and Y are combined to form a 3-, 4-,5-, 6- or 7-membered ring containing from 0 to 2 heteroatoms selectedfrom O, N and S. In a further preferred embodiment, X and Y are combinedto form a 5- or 6-membered ring containing from 0 to 2 heteroatomsselected from O, N and S. In a particularly preferred embodiment, X andY are combined to form a 5- or 6-membered ring containing 0 heteroatoms,1 nitrogen atom or 1 oxygen atom.

[0130] In another preferred embodiment, L² is a bond and Z is —CO₂R¹⁵ or—CO₂NR¹⁵R¹⁶.

[0131] In another group of preferred embodiments, R″ is hydrogen.

[0132] In another group of preferred embodiments, R″ is substituted(C₁-C₈)alkyl. In a preferred embodiment, R″ is (C₁-C₈)alkyl substitutedwith hydroxy, alkylamino (e.g., —NHMe) or carboxy (—CO₂H). In aparticularly preferred embodiment, R″ is (C₃-C₈)alkyl substituted withhydroxy, alkylamino or carboxy.

[0133] In another group of preferred embodiments, R¹ is independentlyhalogen, (C₁-C₄)alkyl, fluoro(C₁-C₄)alkyl, —OR⁵, fluoro(C₁-C₄)alkoxy,—CO₂R⁵, —S(O)_(m)NR⁵R⁶, —S(O)_(m)R⁵ or —CN. In a further preferredembodiment, R¹ is independently halogen or fluoro(C₁-C₄)alkyl. In astill further preferred embodiment, R¹ is halogen or fluoro(C₁-C₄)alkyland the subscript n is 0 or 1. In a particularly preferred embodiment,R¹ is fluoro(C₁-C₄)alkyl and the subscript n is 0 or 1.

[0134] In another group of preferred embodiments, R² is (C₁-C₄)alkyl oraryl.

[0135] In another group of preferred embodiments, R⁴ is hydrogen.

[0136] Also particularly preferred are those embodiments that combinetwo or more of these preferred groups. Accordingly, in one group ofparticularly preferred embodiments, R″ and R⁴ are hydrogen.

[0137] In another group of particularly preferred embodiments, R″ and R⁴are hydrogen and R² is (C₁-C₄)alkyl or aryl.

[0138] In another group of particularly preferred embodiments, R″ and R⁴are hydrogen, R² is independently halogen, (C₁-C₄)alkyl,fluoro(C₁-C₄)alkyl, —OR⁵, fluoro(C₁-C₄)alkoxy, —CO₂R⁵, —S(O),NR⁵R⁶,—S(O)R⁵ or —CN and R² is (C₁-C₄)alkyl or aryl. In a particularlypreferred embodiment, R″ and R⁴ are hydrogen, R¹ is halogen orfluoro(C₁-C₄)alkyl, n is 1 and R² is (C₁-C₄)alkyl or aryl. In a moreparticularly preferred embodiment, R″ and R⁴ are hydrogen, R¹ isfluoro(C₁-C₄)alkyl, n is 1 and R² is (C₁-C₄)alkyl or aryl.

[0139] One group of particularly preferred embodiments is represented bythe formula (IV):

[0140] wherein p, R¹, R², L², X, Y and Z have the meanings and preferredgroupings provided above.

[0141] In a particularly preferred embodiment, X and Y are combined toform a 3-, 4-, 5-, 6- or 7-membered ring containing from 0 to 2heteroatoms selected from O, N and S, L² is a bond and Z is —CO₂R¹ or—CO₂NR¹⁵R¹⁶.

[0142] In another particularly preferred embodiment, X and Y arecombined to form a 3-, 4-, 5-, 6- or 7-membered ring containing from 0to 2 heteroatoms selected from O, N and S and R¹ is independentlyhalogen, (C₁-C₄)alkyl, fluoro(C₁-C₄)alkyl, —OR⁵, fluoro(C₁-C₄)alkoxy,—CO₂R⁵, —S(O)_(m)NR⁵R⁶, —S(O)_(m)R⁵ or —CN.

[0143] In another particularly preferred embodiment embodiment, X and Yare combined to form a 3-, 4-, 5-, 6- or 7-membered ring containing from0 to 2 heteroatoms selected from O, N and S and R² is (C₁-C₄)alkyl oraryl.

[0144] In another particularly preferred embodiment embodiment, X and Yare combined to form a 3-, 4-, 5-, 6- or 7-membered ring containing from0 to 2 heteroatoms selected from O, N and S, R¹ is independentlyhalogen, (C₁-C₄)alkyl, fluoro(C₁-C₄)alkyl, —OR⁵, fluoro(C₁-C₄)alkoxy,—CO₂R⁵, —S(O)_(m)NR⁵R⁶, —S(O)_(m)R⁵ or —CN and R² is (C₁-C₄)alkyl oraryl.

[0145] In another particularly preferred embodiment embodiment, R¹ isindependently halogen, (C₁-C₄)alkyl, fluoro(C₁-C₄)alkyl, —OR⁵,fluoro(C₁-C₄)alkoxy, —CO₂R⁵, —S(O)_(m)NR⁵R⁶, —S(O)_(m)R⁵ or —CN and R²is (C₁-C₄)alkyl or aryl.

[0146] In another particularly preferred embodiment embodiment, R¹ isindependently halogen, (C₁-C₄)alkyl, fluoro(C₁-C₄)alkyl, —OR⁵,fluoro(C₁-C₄)alkoxy, —CO₂R⁵, —S(O)_(m)NR⁵R⁶, —S(O)_(m)R⁵ or —CN, L² is abond and Z is —CO₂R¹⁵ or —CO₂NR¹⁵R¹⁶.

[0147] In another particularly preferred embodiment embodiment, R² is(C₁-C₄)alkyl or aryl, L² is a bond and Z is —CO₂R¹⁵ or —CO₂NR¹⁵R¹⁶.

[0148] In another particularly preferred embodiment embodiment, R¹ isindependently halogen, (C₁-C₄)alkyl, fluoro(C₁-C₄)alkyl, —OR⁵,fluoro(C₁-C₄)alkoxy, —CO₂R⁵, —S(O)_(m)NR⁵R⁶, —S(O)_(m)R⁵ or N, R² is(C₁-C₄)alkyl or aryl, L² is a bond and Z is —CO₂R¹⁵ or —CO₂NR¹⁵R¹⁶.

[0149] In a particular embodiment, the present invention provides thefollowing compounds:

[0150] In further particular embodiments, the present invention providespharmaceutically acceptable salts of the above compounds. For example,in a certain embodiment the present invention provides benzenesulfonicacid salts of the above compounds.

[0151] Compositions

[0152] In another aspect, the present invention provides pharmaceuticalcompositions comprising one or more compounds of the invention incombination with a diagnostically or pharmaceutically acceptable carrieror excipient. The subject compositions are useful for treating orpreventing conditions and disorders mediated by MCHR, such as obesityand eating disorders, e.g., anorexia nervosa. The compounds of thepresent invention can be prepared and administered in a wide variety oforal and parenteral dosage forms. Thus, the compounds of the presentinvention can be administered by injection, for example, intravenously,intramuscularly, intracutaneously, subcutaneously, intraduodenally orintraperitoneally. Also, the compounds described herein can beadministered by inhalation, for example, intranasally. Additionally, thecompounds of the present invention can be administered transdermally.Other routes of administration are also contemplated for use with thecompounds of the present invention, including depot administration andrectal administration.

[0153] For preparing pharmaceutical compositions from the compounds ofthe present invention, pharmaceutically acceptable carriers can beeither solid or liquid. Solid form preparations include powders,tablets, pills, capsules, cachets, suppositories, and dispersiblegranules. A solid carrier can be one or more substances which may alsoact as diluents, flavoring agents, binders, preservatives, tabletdisintegrating agents, or an encapsulating material.

[0154] In powders, the carrier is a finely divided solid which is in amixture with the finely divided active component. In tablets, the activecomponent is mixed with the carrier having the necessary bindingproperties in suitable proportions and compacted in the shape and sizedesired.

[0155] The powders and tablets preferably contain from about 5% or 10%to 70% of the active compound. Suitable carriers are magnesiumcarbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin,starch, gelatin, tragacanth, methylcellulose, sodiumcarboxymethylcellulose, a low melting wax, cocoa butter, and the like.The term “preparation” is intended to include the formulation of theactive compound with encapsulating material as a carrier providing acapsule in which the active component with or without other carriers, issurrounded by a carrier, which is thus in association with it.Similarly, cachets and lozenges are included. Tablets, powders,capsules, pills, cachets, and lozenges can be used as solid dosage formssuitable for oral administration.

[0156] For preparing suppositories, a low melting wax, such as a mixtureof fatty acid glycerides or cocoa butter, is first melted and the activecomponent is dispersed homogeneously therein, as by stirring. The moltenhomogeneous mixture is then poured into convenient sized molds, allowedto cool, and thereby to solidify.

[0157] Liquid form preparations include solutions, suspensions andemulsions, for example, water or water/propylene glycol solutions. Forparenteral injection, liquid preparations can be formulated in solutionin aqueous polyethylene glycol solution.

[0158] Aqueous solutions suitable for oral use can be prepared bydissolving the active component in water and adding suitable colorants,flavors, stabilizers, and thickening agents as desired. Aqueoussuspensions suitable for oral use can be made by dispersing the finelydivided active component in water with viscous material, such as naturalor synthetic gums, resins, methylcellulose, sodiumcarboxymethylcellulose, and other well-known suspending agents.

[0159] Also included are solid form preparations which are intended tobe converted, shortly before use, to liquid form preparations for oraladministration. Such liquid forms include solutions, suspensions andemulsions. These preparations may contain, in addition to the activecomponent, colorants, flavors, stabilizers, buffers, artificial andnatural sweeteners, dispersants, thickeners, solubilizing agents and thelike.

[0160] The pharmaceutical preparation is preferably in unit dosage form.In such form the preparation is subdivided into unit doses containingappropriate quantities of the active component. The unit dosage form canbe a packaged preparation, the package containing discrete quantities ofpreparation, such as packeted tablets, capsules and powders in vials orampoules. Also, the unit dosage form can be a capsule, tablet, cachet,or lozenge itself, or it can be the appropriate number of any of thesein packaged form.

[0161] The quantity of active component in a unit dose preparation maybe varied or adjusted from 0.1 mg to 1000 mg, preferably 1.0 mg to 100mg according to the particular application and the potency of the activecomponent. The composition can, if desired, also contain othercompatible therapeutic agents.

[0162] In therapeutic use for the treatment or prevention of conditionsand disorders associated with MCHR, the compounds utilized in thepharmaceutical method of the invention are administered at the initialdosage of about 0.001 mg/kg to about 100 mg/kg daily. A daily dose rangeof about 0.1 mg/kg to about 10 mg/kg is preferred. The dosages, however,may be varied depending upon the requirements of the patient, theseverity of the condition being treated and the compound being employed.Determination of the proper dosage for a particular situation is withinthe skill of the practitioner. Generally, treatment is initiated withsmaller dosages which are less than the optimum dose of the compound.Thereafter, the dosage is increased by small increments until theoptimum effect under the circumstances is reached. For convenience, thetotal daily dosage may be divided and administered in portions duringthe day, if desired.

[0163] The compositions may be advantageously combined and/or used incombination with agents useful in the treatment and/or prevention ofobesity and eating disorders and pathologies associated therewith (e.g.,cardiovascular disease and hypertension). In many instances,administration of the subject compounds or compositions in conjunctionwith these alternative agents enhances the efficacy of such agents.Accordingly, in some instances, the present compounds, when combined oradministered in combination with, e.g., anti-obesity agents, can be usedin dosages which are less than the expected amounts when used alone, orless than the calculated amounts for combination therapy.

[0164] Suitable agents for combination therapy include those that arecurrently commercially available and those that are in development orwill be developed. Exemplary agents useful in the treatment of obesityinclude β₃ adrenergic receptor agonists, leptin or derivatives thereofand neuropeptide Y antagonists. Exemplary agents useful in the treatmentof anxiety and/or mood disorders include benzodiazepines, e.g.,alprazolam, chlordiazepoxide, clonazepam, clorazepate, diazepam,lorazepam, oxazepam, and the like; heterocyclic antidepressants, e.g,amitriptyline, nortriptyline, imipramine, desipramine, doxepin,trimipramine, clomipramine, protryptyline, amoxapine and maprotiline;monoaamine oxidase inhibitors (MAOIs), e.g., phenelzine andtranylcypromine; serotonin reuptake inhibitors (SRIs); selectiveserotonin reuptake inhibitors (SSRIs), e.g., fluoxetine, fluvoxamine,paroxetine and sertraline; serotonergic-noradrenergic antidepressants,e.g., venlafaxine; 5-HT2 antagonists, e.g., trazadone, nefazodone andmirtazapine; and catecholaminergic antidepressants, e.g., buproprion.

[0165] Methods of Use

[0166] In yet another aspect, the present invention provides methods ofusing one or more compounds of the invention to treat or prevent acondition or disorder associated with eating behavior, energyhomeostasis or anxiety. Exemplary conditions and disorders associatedwith eating behavior, energy homeostasis and anxiety include eatingdisorders, such as anorexia nervosa and bulimia, obesity, anxietydisorders, e.g., generalized anxiety disorder, panic attacks, panicdisorder and obsessive-compulsive disorder (OCD), and mood disorders,e.g., depression and bipolar disorders. Methods of using a compound ofthe invention to treat or prevent a condition or disorder associatedwith eating behavior include methods of modifying eating behavior orfood intake, for example, stimulating or suppressing eating behavior orincreasing or decreasing food intake. The methods comprise administeringto a subject in need thereof a therapeutically effective amount of acompound of the invention.

[0167] In another aspect, the present invention provides methods ofusing a compound of the invention to treat or prevent a condition ordisorder mediated by MCHR. The methods comprise administering to asubject in need thereof a therapeutically effective amount of a compoundof the invention.

[0168] In still another aspect, the present invention provides methodsof using a compound of the invention to modulate MCHR. The methodscomprise contacting a cell with a compound of the invention.

[0169] The compounds of the invention may also modulate G-proteincoupled receptors related to MCHR, e.g., MCHR2 (see InternationalPublication Nos. WO 00/49046 and WO 01/07606).

[0170] Preparation of the Compounds

[0171] The present invention provides a process for the preparation of acompound of formula I.

[0172] A general synthetic route is depicted in Scheme 1, which outlinesthe condensation of substituted aryl moiety A, with a bicyclic structureB to produce a compound of formula C, wherein the variables are asdefined as above. In formula A, D¹ is hydrogen, halogen, —C(O)R⁷, —CO₂R⁸or —C(O)NR⁵R⁶, wherein R⁵, R⁶, R⁷ and R⁸ are defined as above, and D² isa bond, —N(R″)—, —N(protecting group)-, —S— or —O—, wherein R″ isdefined as above and protecting group is an amino protecting group.Conventional amino protecting groups consist of known groups which areused to protectively block an amino group during the synthesisprocedures described herein. These conventional blocking groups arereadily removable, i.e., they can be removed, if desired, by procedureswhich will not cause cleavage or other disruption of the remainingportions of the molecule. Suitable protecting groups for the compoundsof the present invention will be recognized from the present applicationtaking into account the level of skill in the art, and with reference tostandard textbooks, such as Greene, T. W. et al,. Protective Groups inOrganic Synthesis, Wiley, New York (1991).

[0173] In formula B, E¹ is hydrogen, —C(O)R⁷, —CO₂R⁸ or —C(O)NR⁵R⁶,wherein R⁵, R⁶, R⁷ and R⁸ are defined as above, and E² is ═O or —NR⁵R⁶,wherein R⁵ and R⁶ are defined as above. When a compound of formula A,wherein D¹ is hydrogen and D² is —N(R″)— or —N(protecting group)-, —S—,or —O—, reacts with a compound of formula B, wherein E¹ is hydrogen andE² is ═O or a protected version thereof (e.g., an acetal), under thetypical Fisher indolization conditions, a compound of formula C isproduced.

[0174] One of skill in the art will understand that the synthesisprovided above can be modified to use different starting materials andalternate reagents to accomplish the desired transformations. Forexample, a compound of formula A, wherein D¹ is a leaving group such asCl, Br, I or toluenesulfonate, can react with a compound of formula B,wherein E² is ═O or a protected version thereof, via apalladium-catalyzed coupling reaction to produce a compound of formulaC. Also, a compound of formula A, wherein D¹ is a leaving group and D²is a nitro group, can react with a compound of formula B, wherein E¹ isCO₂R and wherein E² is ═O or a protected version thereof, to produce acompound of formula C. Accordingly, the synthesis and reagents describedherein are all expressed as non-limiting embodiments.

[0175] Materials represented by formula A are available commercially(Aldrich Chemical), or can be obtained synthetically followingliterature procedures.

[0176] One way to prepare compounds represented by formula B is by theRobinson annulation process between a cyclic ketone and a substitutedenone followed by saturation of the double bond. One of the skill in theart will readily appreciate that other methods are available. Therelative stereochemistry and absolute stereochemistry can be controlledin the process. The individual forms of compounds of formula B, e.g.,diastereomers and enantiomers, can be formed by stereocontrolledreactions, or may be separated, e.g., by chromatographic techniques(diastereomers) and by resolution (enantiomers).

[0177] Analysis of the Compounds The activity of MCHR polypeptides canbe assessed using a variety of in vitro and in vivo assays to determinefunctional, chemical and physical effects, e.g., measuring ligandbinding (e.g., radioactive ligand binding), second messenger (e.g.,cAMP, cGMP, IP₃, DAG, or Ca²⁺) levels, ion flux, phosphorylation levels,transcription levels, neurotransmitter levels, and the like.Furthermore, such assays can be used to test for antagonists andagonists of MCHR. Screening assays may be used to identify modulatorsthat can be used as therapeutic agents, e.g., antagonists of MCHRactivity.

[0178] Modulators of MCHR activity can be tested using MCHR polypeptidesas described above, either recombinant or naturally occurring (e.g.,endogenous). The protein can be isolated, expressed in a cell, expressedin a membrane derived from a cell, expressed in tissue or in an animal,either recombinant or naturally occurring. For example, kidney cells,liver cells, colon cells, transformed cells, or membranes can be used.Modulation is tested using one of the in vitro or in vivo assaysdescribed herein. Signal transduction can also be examined in vitro withsoluble or solid state reactions, using a chimeric molecule such as anextracellular domain of a receptor covalently linked to a heterologoussignal transduction domain, or a heterologous extracellular domaincovalently linked to the transmembrane and or cytoplasmic domain of areceptor. Gene amplification can also be examined. Furthermore,ligand-binding domains of the protein of interest can be used in vitroin soluble or solid state reactions to assay for ligand binding.

[0179] Ligand binding to MCHR, a domain, or chimeric protein can betested in solution, in a bilayer membrane, attached to a solid phase, ina lipid monolayer, or in vesicles. Binding of a modulator can be testedusing, e.g., changes in spectroscopic characteristics (e.g.,fluorescence, absorbance, refractive index) hydrodynamic (e.g., shape),chromatographic, or solubility properties.

[0180] MCHR-G-protein interactions can also be examined, by, forexample, analysis of binding of the G-protein to MCHR or its releasefrom MCHR can be examined. For example, in the absence of GTP, anactivator will lead to the formation of a tight complex of a G protein(all three subunits) with MCHR. This complex can be detected in avariety of ways, as noted above. Such an assay can be modified to searchfor antagonists. In one embodiment, an activator is added to MCHR and Gprotein in the absence of GTP, allowed to form a tight complex, and thenscreened for antagonists by looking at dissociation of the MCHR-Gprotein complex. In the presence of GTP, release of the alpha subunit ofthe G protein from the other two G protein subunits serves as acriterion of activation.

[0181] An activated or inhibited G-protein will in turn alter theproperties of downstream effectors such as proteins, enzymes andchannels. The classic examples are the activation of cGMPphosphodiesterase by transducin in the visual system, adenylate cyclaseby the stimulatory G-protein, phospholipase C by Gq and other cognate Gproteins, and modulation of diverse channels by Gi and other G proteins.Downstream consequences can also be examined such as generation ofdiacyl glycerol and IP3 by phospholipase C, and in turn, for calciummobilization by IP3.

[0182] Activated MCHR becomes a substrate for kinases that phosphorylatethe C-terminal tail of the receptor (and possibly other sites as well).Thus, activators will promote the transfer of ³²P from gamma-labeled ATPto the receptor, which can be assayed with a scintillation counter. Thephosphorylation of the C-terminal tail will promote the binding ofarrestin-like proteins and will interfere with the binding ofG-proteins. The kinase/arrestin pathway plays a key role in thedesensitization of many GPCR receptors. For a general review of GPCRsignal transduction and methods of assaying signal transduction, see,e.g., Methods in Enzymology, vols. 237 and 238 (1994) and volume 96(1983); Bourne et al., Nature 10:349:117-27 (1991); Bourne et al.,Nature 348:125-32 (1990); Pitcher et al., Annu. Rev. Biochem. 67:653-92(1998).

[0183] Samples or assays that are treated with a potential MCHRantagonist or agonist are compared to control samples without the testcompound, to examine the extent of modulation. Control samples(untreated with agonists or antagonist) are assigned a relative MCHRactivity value of 100. Inhibition of MCHR is achieved when the MCHRactivity value relative to the control is about 90%, optionally 50%,optionally 25-0%. Activation of MCHR is achieved when the MCHR activityvalue relative to the control is 110%, optionally 150%, 200-500%, or1000-2000%.

[0184] Changes in ion flux may be assessed by determining changes inpolarization (i.e., electrical potential) of the cell or membraneexpressing MCHR. One means to determine changes in cellular polarizationis by measuring changes in current (thereby measuring changes inpolarization) with voltage-clamp and patch-clamp techniques, e.g., the“cell-attached” mode, the “inside-out” mode, and the “whole cell” mode(see, e.g., Ackerman et al., New Engl. J. Med. 336:1575-1595 (1997)).Whole cell currents are conveniently determined using the standardmethodology (see, e.g., Hamil et al., PFlugers. Archiv. 391:85 (1981).Other known assays include radiolabeled ion flux assays and fluorescenceassays using voltage-sensitive dyes (see, e.g., Vestergarrd-Bogind etal., J. Membrane Biol. 88:67-75 (1988); Gonzales & Tsien, Chem. Biol.4:269-277 (1997); Daniel et al., J. Pharmacol. Meth. 25:185-193 (1991);Holevinsky et al., J. Membrane Biology 137:59-70 (1994)). Generally, thecompounds to be tested are present in the range from 1 pM to 100 mM.

[0185] The effects of the test compounds upon the function of thepolypeptides can be measured by examining any of the parametersdescribed above. Any suitable physiological change that affects MCHRactivity can be used to assess the influence of a test compound on thepolypeptides of this invention. When the functional consequences aredetermined using intact cells or animals, one can also measure a varietyof effects such as transmitter release, hormone release, transcriptionalchanges to both known and uncharacterized genetic markers (e.g.,northern blots), changes in cell metabolism such as cell growth or pHchanges, and changes in intracellular second messengers such as Ca²⁺,IP3 or cAMP.

[0186] Preferred assays for MCHR include cells that are loaded with ion-or voltage-sensitive dyes to report receptor activity. Assays fordetermining activity of such receptors can also use known agonists andantagonists for other G-protein coupled receptors as negative orpositive controls to assess activity of tested compounds. In assays foridentifying modulatory compounds (e.g., agonists, antagonists), changesin the level of ions in the cytoplasm or membrane voltage will bemonitored using an ion-sensitive or membrane voltage fluorescentindicator, respectively. Among the ion-sensitive indicators and voltageprobes that may be employed are those disclosed in the Molecular Probes1997 Catalog. For G-protein coupled receptors, promiscuous G-proteinssuch as Gα15 and Gα16 can be used in the assay of choice (Willie et al.,Proc. Natl Acad. Sci. USA 88:10049-10053 (1991)). Such promiscuousG-proteins allow coupling of a wide range of receptors to signaltransduction pathways in heterologous cells.

[0187] Receptor activation typically initiates subsequent intracellularevents, e.g., increases in second messengers such as IP3, which releasesintracellular stores of calcium ions. Activation of some G-proteincoupled receptors stimulates the formation of inositol triphosphate(IP3) through phospholipase C-mediated hydrolysis ofphosphatidylinositol (Berridge & Irvine, Nature 312:315-21 (1984)). IP3in turn stimulates the release of intracellular calcium ion stores.Thus, a change in cytoplasmic calcium ion levels, or a change in secondmessenger levels such as IP3 can be used to assess G-protein coupledreceptor function. Cells expressing such G-protein coupled receptors mayexhibit increased cytoplasmic calcium levels as a result of contributionfrom both intracellular stores and via activation of ion channels, inwhich case it may be desirable although not necessary to conduct suchassays in calcium-free buffer, optionally supplemented with a chelatingagent such as EGTA, to distinguish fluorescence response resulting fromcalcium release from internal stores.

[0188] Other assays can involve determining the activity of receptorswhich, when activated, result in a change in the level of intracellularcyclic nucleotides, e.g., cAMP or cGMP, by activating or inhibitingdownstream effectors such as adenylate cyclase. There are cyclicnucleotide-gated ion channels, e.g., rod photoreceptor cell channels andolfactory neuron channels that are permeable to cations upon activationby binding of cAMP or cGMP (see, e.g., Altenhofen et al., Proc. Natl.Acad. Sci. U.S.A. 88:9868-9872 (1991) and Dhallan et al., Nature347:184-187 (1990)). In cases where activation of the receptor resultsin a decrease in cyclic nucleotide levels, it may be preferable toexpose the cells to agents that increase intracellular cyclic nucleotidelevels, e.g., forskolin, prior to adding a receptor-activating compoundto the cells in the assay. Cells for this type of assay can be made byco-transfection of a host cell with DNA encoding a cyclicnucleotide-gated ion channel, GPCR phosphatase and DNA encoding areceptor (e.g., certain glutamate receptors, muscarinic acetylcholinereceptors, dopamine receptors, serotonin receptors, and the like),which, when activated, causes a change in cyclic nucleotide levels inthe cytoplasm.

[0189] In one embodiment, changes in intracellular cAMP or cGMP can bemeasured using immunoassays. The method described in Offermanns & Simon,J. Biol. Chem. 270:15175-15180 (1995) may be used to determine the levelof cAMP. Also, the method described in Felley-Bosco et al., Am. J. Resp.Cell and Mol. Biol. 11:159-164 (1994) may be used to determine the levelof cGMP. Further, an assay kit for measuring cAMP and/or cGMP isdescribed in U.S. Pat. No. 4,115,538, herein incorporated by reference.In another embodiment, phosphatidyl inositol (PI) hydrolysis can beanalyzed according to U.S. Pat. No. 5,436,128, herein incorporated byreference.

[0190] In another embodiment, transcription levels can be measured toassess the effects of a test compound on signal transduction. A hostcell containing the protein of interest is contacted with a testcompound for a sufficient time to effect any interactions, and then thelevel of gene expression is measured. The amount of time to effect suchinteractions may be empirically determined, such as by running a timecourse and measuring the level of transcription as a function of time.The amount of transcription may be measured by using any method known tothose of skill in the art to be suitable. For example, mRNA expressionof the protein of interest may be detected using northern blots or theirpolypeptide products may be identified using immunoassays.Alternatively, transcription based assays using a reporter gene may beused as described in U.S. Pat. No. 5,436,128, herein incorporated byreference. The reporter genes can be, e.g., chloramphenicolacetyltransferase, firefly luciferase, bacterial luciferase,β-galactosidase and alkaline phosphatase. Furthermore, the protein ofinterest can be used as an indirect reporter via attachment to a secondreporter such as green fluorescent protein (see, e.g., Mistili &Spector, Nature Biotechnology 15:961-964 (1997)).

[0191] The amount of transcription is then compared to the amount oftranscription in either the same cell in the absence of the testcompound, or it may be compared with the amount of transcription in asubstantially identical cell that lacks the protein of interest. Asubstantially identical cell may be derived from the same cells fromwhich the recombinant (or non-recombinant) cell line was prepared butwhich had not been modified by introduction of heterologous DNA. Anydifference in the amount of transcription indicates that the testcompound has in some manner altered the activity of the protein ofinterest.

[0192] The following examples are provided by way of illustration onlyand not by way of limitation. Those of skill in the art will readilyrecognize a variety of noncritical parameters that could be changed ormodified to yield essentially similar results.

EXAMPLES

[0193] Reagents and solvents used below can be obtained from commercialsources such as Aldrich Chemical Co. (Milwaukee, Wis., USA). ¹H-NMRspectra were recorded on a Varian Gemini 400 MHz NMR spectrometer.Significant peaks are tabulated in the order: multiplicity (s, singlet;d, doublet; t, triplet; q, quartet; m, multiplet; br s, broad singlet),coupling constant(s) in Hertz (Hz) and number of protons. ElectronIonization (El) mass spectra were recorded on a Hewlett Packard 5989Amass spectrometer. Mass spectrometry results are reported as the ratioof mass over charge, followed by the relative abundance of each ion (inparentheses). A single m/e value is reported for the M+H (or, as noted,M−H) ion containing the most common atomic isotopes. Isotope patternscorrespond to the expected formula in all cases. Electrospray ionization(ESI) mass spectrometry analysis was conducted on a Hewlett-Packard 1100MSD electrospray mass spectrometer using the HP1 100 HPLC for sampledelivery. Normally the analyte was dissolved in methanol at 0.1 mg/mLand 1 microliter was infused with the delivery solvent into the massspectrometer, which scanned from 100 to 1500 daltons. All compoundscould be analyzed in the positive ESI mode, using 1:1 acetonitrile/waterwith 1% acetic acid as the delivery solvent. The compounds providedbelow could also be analyzed in the negative ESI mode, using 2 mM NH₄OAcin acetonitrile/water as delivery solvent. Analytical HPLC analysis wasconducted on a Hewlett-Packard Series 1050 system equipped with a C18reverse phase column (4.6 mm×150 mm) manufactured by Shiseido Co.,Japan. Gradient elution was performed using variable percentage ofacetonitrile and water (each with 0.1 % trifluoroacetic acid added) as amobile phase. Optical purity analysis was also conducted on aHewlett-Packard Series 1050 system equipped with a chiral HLPC column(ChiralPak AD, 4.6 mm×150 mm) purchased from Chiral Technology.Isopropanol (3%) and hexane (97%) containing 0.1% diethylarnine was usedas a mobile phase.

Example 1

[0194]

[0195] Compound 4 was prepared in 4 steps, as follows.

[0196] Step 1. Robinson Annulation. A mixture of N-benzyl-4-piperidone(500 g, 2.65 mol) and pyrrolidine (330 mL, d 0.852, 3.96 mol) in toluene(2 L) was heated at refluxing while water from elimination was removedwith a Dean-Stark trap. After 8 h, 70 mL of water were collected, andthe volume of collected water ceased to increase further. GC analysisrevealed the presence of the starting N-benzyl-4-piperidone and productenamine. The solvent and excess pyrrolidine were evaporated underreduced pressure (vacuum, 60 torr; heating bath, 50° C.). The residuewas dissolved in 500 mL of toluene, and evaporated again to give a darkoil (630 g).

[0197] The resulting enamine was dissolved in anhydrous dioxane (2 L)and filtered into a 5-L three-necked flask equipped with a mechanicstirrer, a condenser and an addition funnel. 3-Penten-2-one (333 g, 2.78mol) was added to the reaction vessel in 20 min. The reaction mixturewas heated to reflux for 25 h. After cooled to near r.t., NaOMe (6.7 g,0.125 mol) was added and the mixture was heated to reflux for 6 h. Apremixed solution of AcONa (200 g) in 400 mL water and glacial AcOH (400mL) was added to the reaction mixture after cooling to near roomtemperature. The reaction mixture was heated to reflux for an additional5 h. Approximately 1 L of solvent (and possibly pyrrolidine) wasdistilled out, the rest of the reaction mixture was cooled to r.t,brought to slightly basic (pH 8-9) with 2 N NaOH (2.5 L). After layerseparation, aqueous phase was extracted with AcOEt (3 L). The organicextracts were combined, washed with brine, and filtered through a shortsilica gel plug to give a dark clear solution. The filtrate wasconcentrated under reduced pressure to a thick oil (670 g). Thismaterial was used in the resolution step directly.

[0198] Step 2. Resolution

[0199] To a stirred hot solution of the racemic isoquinolinone free base(626 g, 2.45 mol) in 95% ethanol (800 mL) was added solution ofdi-O-p-toluoyl-L-tartaric acid (945 g, 2.45 mol) in hot ethanol (1500mL). Precipitation of the less soluble diasteromeric salt occurredgenerally as soon of the mixing was completed. The mixture was heated ina hot water bath (80° C.) with gentle stirring for 1 h and allowed tocool to r.t. slowly (typically overnight). The precipitate was collectedby filtration and rinsed wih cold 95% ethanol (800 mL). The solid(off-white) was triturated in hot 95% ethanol (1500 mL) and collectedafter cooling (typically after standing at r.t. overnight) by filtrationand washed with cold ethanol. An off-white solid (340 g, ca. 98% ee) wasobtained after two triturations.

[0200] Compound 2a was liberated from the salt by neutralization withNaOH and extraction with AcOEt.

[0201] A flask for the Parr shaker hydrogenation apparatus was chargedwith the N-benzylisoqinolinone compound from previous step (120 g, 0.47mol), 10% Pd/C (12 g, contains 50% water), di-t-butyl dicarbonate (133g, 0.61 mol), and ethanol (1200 mL, 200 proof). The reactions took placeunder a hydrogen pressure of 60 psi. The hydrogen presure droppedquickly in the first 2 h, frequent recharges were needed. The reactionwas typically left to go undisturbed for 8 h or longer. No further H₂consumption was observed. The reaction mixture was filtered through aCelite pad, rinsed with ethanol. Filtrate was concentrated to theproduct as a thick oil, which solidified on standing to give a whitesolid. This material was used in the next step without furtherpurification.

[0202] The solution of the N-Boc-isoquinolinone compound from previousstep in propanol (200 mL) was placed in a pressure resistant vessel.Concentrated H₂SO₄ (13 mL) was added slowly. Gas release took placeimmediately and subsided after 45 min. 4-Trifluorophenylhydrazine (16.56g, 94.0 mmol) was added. The mixture was stirred for 1 h at r.t followedby 3 h at refluxing or until gas evolution stopped. At this time, MSanalysis indicted hydrozone as the major component of the reactionmixture. The reaction vessel was capped and heated at 90° C. for 36 huntil the completion of reaction, as monitored by TLC (10: 1:0.1,CH₂Cl₂/MeOH/NH₄OH), and ES-MS in positive mode. At the completion of thereaction, the reaction mixture was poured to a stirred solution of 1 NNaOH (some precipitates were formed). The mixture was extracted withdichloromethane three times. The combined organic extracts were washedwith water, dried over NaSO₄, filtered, and concentrated to give asolid. The residue was triturated with CH₂Cl₂. The solid was collectedby filtration. The product could be purified by chromatography on silicagel column with a gradient elution of increasing polarity from 20:1:0.1to 6:1:0.1 CH₂Cl₂/MeOH/NH₄OH to obtain compound 4 as the major product.¹H NMR δ 11.2 (s, 1H), 7.80 (s, 1H), 7.43 (d, J=5.4 Hz, 1H), 7.28 (d,J=5.4 Hz, 1H), 3.40 (d, J=6.0 Hz, 1H), 3.01 (d, J=8.0 Hz, 1H), 2.75 (d,J=10 Hz, 1H), 2.60 (m, 2H), 2.40 (m, 2H), 1.82 (d, J=8 Hz, 1H), 1.56 (m,1H), 1.38 (d, J=5.4 Hz, 3H), 1.25 (m, 2H). MS (ES): 309 [M+H]⁺.

Example 2

[0203]

[0204] Compound 5 was synthesized in 3 steps, as follows.

[0205] Step 1. To a 500 mL flask containing iPr₂NH (16.82 mL, 120 mmol)in THF (200 mL) at −78° C. was added n-BuLi (48 mL, 2.5 M/hexanes, 120mmol). After stirring for 30 min at −78° C.,4-Tetrahydro-pyran-4-carboxylic acid methyl ester (11.86 mL, 100 mmol)was added. After stirring for an additional 45 min, HMPA (10 mL) andallyl iodide (11.9 mL, 130 mmol) were added. The reaction was maintainedfor 20 min at the low temperature and allowed to warm up to r.t. Thereaction mixture was poured into water and extracted with ether. Theorganic layer was washed with brine, dried with anhydrous Na₂SO₄,concentrated by rotary evaporation and purified by flash chromatographyon silica gel with a gradient elution of 20-30% EtOAc/hexanes to yield aclear oil (16.6 g).

[0206] Step 2. The above alkylation product (15.26 g, 83 mmol) wasstirred with NaIO₄ (39.0 g, 182 mmol) and OsO₄ (70 mg) in iPrOH (400 mL)and H₂O (400 mL) overnight. The reaction mixture was poured into waterand extracted with EtOAc. The organic layer was separated, washed withbrine, dried with anhydrous Na₂SO₄, concentrated by rotary evaporationand purified by flash chromatography on silica gel with a gradientelution of 40-70% EtOAc/hexanes to yield the product,4-(2-oxo-ethyl)-tetrahydropyran-4-carboxylic acid methyl ester, as anoil (8.8 g).

[0207] Step 3. The above aldehyde (1.86 g, 10 mmol) was stirred withamine 4 (3.08 g, 10 mmol) and NaBH(OAc)₃ (8.48 g, 40 mmol) in ClCH₂CH₂Cl(50 mL) overnight. It was poured into a dilute aq. ammonia solution andextracted with EtOAc. The organic layer was separated, washed withbrine, dried with anhydrous Na₂SO₄, concentrated by rotary evaporationand purified by flash chromatography on silica gel with a gradientelution of EtOAc, 10% MeOH/EtOAc, 10% MeOH/CH₂Cl₂, 20% MeOH/CH₂Cl₂ and30% MeOH/CH₂Cl₂ to yield 5 as a solid (3.0 g). ¹H NMR δ (DMSO, 400 MHz),11.20 (s, 1H), 7.88 (s, 1H), 7.44 (d, J=8.5 Hz, 1H), 7.30 (d, J=8.5 Hz,1H), 3.73 (m, 1H), 3.68 (s, 3H), 3.33 (m, 2H), 3.22 (m, 1H), 2.88 (m,1H), 2.75 (m, 1H), 2.60 (m, 1H), 2.42 (m, 1H), 2.26 (m, 2H), 1.98 (m,2H), 1.7-1.95 (m, 5H), 1.51 (m, 2H), 1.38 (d, J=6.5 Hz, 3H), 1.39 (m,1H), 1.20 (m, 1H). MS (ES): 479 [M+H].

Example 3

[0208]

[0209] A sample of compound 5 (0.080 g, 0.17 mmol) in TBF (2 mL) wasreduced with LiAlH₄ (0.400 mL, 1M/THF, 0.40 mmol) in TBF (2 mL). At thecompletion of the reduction, the reaction was quenched with an aqueoussolution of 10% Na₂SO₄. The precipitate was removed by filtration, andthe organic filtrate was washed with brine, dried with anhydrous Na₂SO₄,concentrated by rotary evaporation. Purification by flash chromatographyon silica gel with a gradient elution of 5-30% MeOH/CH₂Cl₂ to yield 6 asa white solid (0.065 g). ¹H NMR δ (DMSO, 400 MHz), 11.22 (s, 1 H), 7.79(s, 1 H), 7.44 (d, J=8.5 Hz, 1 H), 7.29 (d, J=8.5 Hz, 1 H), 5.35 (s, 1H), 3.56 (m, 4 H), 3.31 (m, 3 H), 3.02 (m, 1 H), 2.74 (m, 1 H), 2.62 (m,1 H), 2.43 (m, 3 H), 1.95 (m, 1 H), 1.86 (m, 1 H), 1.75 (m, 1 H), 1.60(m, 2 H), 1.25-1.5 (m, 10 H). MS (ES): 451 [M+H].

Example 4

[0210]

[0211] A mixture of ester 5 (0.80 g, 1.7 mmol) and LiOH.H₂O (0.80 g, 19mmol) in dioxane (10 mL) and water (5 mL) was heated at refluxing for 7h. The reaction mixture was cooled. On acidification with HOAc (to pHca. 5) white precipitate was formed. The solid was collected byfiltration, rinsed with water and finally with ether to yield thecompound 7 as white solid (0.50 g). ¹H NMR δ (DMSO, 500 MHz), 11.21 (s,1 H), 7.79 (s, 1 H), 7.44 (d, J=8.5 Hz, 1 H), 7.29 (d, J=8.5 Hz, 1 H),3.71 (m, 2 H), 3.31 (m, 2 H), 2.97 (m, 1 H), 2.75 (m, 1 H), 2.62 (m, 1H), 2.44 (m, 4 H), 1.6-2.05 (m, 8 H), 1.2-1.5 (m, 8 H). MS (ES): 465[M+H].

Example 5

[0212]

[0213] To a mixture of acid 7 (1.86 g, 4 mmol) in CH₂Cl₂ (60 mL) with 4drops of DMF was added oxalyl chloride (17 mL, 2M in CH₂Cl₂, 34 mmol).After stirring at r.t for 1 h, the mixture was concentrated and pumpedunder high vacuum to obtain a solid. To this solid was added sat.solution of NH₄OH in CH₂Cl₂ (60 mL). The mixture was stirred overnightand directly loaded onto a chromatographic column with a gradientelution of 10-20% MeOH/CH₂Cl₂ with increasing percentage (0-10%) ofNH₄OH added to yield compound 8 as a white solid (1.674 g). ¹H NMR δ(DMSO, 400 MHz), 11.28 (s, 1H), 7.80 (s, 1H), 7.45 (d, J=8.5 Hz, 1H),7.29 (d, J=8.5 Hz, 1H), 7.00 (s, br, 2H), 3.67 (m, 2H), 3.38 (m, 5H),2.6-3.0 (m, 3H), 2.43 (m, 1H), 2.20 (m, 1H), 1.99 (m, 3H), 1.6-1.9 (m,4H), 1.35-1.5 (m, 7H). MS (ES): 464 [M+H].

Example 6

[0214]

[0215] To a mixture of acid 7 (0.120 g, 0.256 mmol), DMF (2 drops) inCH₂Cl₂ (2 mL) was added (COCl)₂ (1.2 mL, 2M/CH₂Cl₂, 2.4 mmol). When thegas release ceased, the mixture was placed under high vacuum to obtain asolid. This solid was resuspended in CH₂Cl₂ (2 mL), to it was added(S)(+) 2-amino-1-propanol (0.100 mL, 1.7 mmol) and NEt₃ (0.140 mL, 1mmol). After 1 h stirring at r.t, the mixture was poured into saturatedsolution of NaHCO₃ and extracted with EtOAc. The organic layer wasseparated, washed with brine, dried with anhydrous Na₂SO₄, concentratedby rotary evaporation and purified by flash chromatography on silica gelwith a gradient elution of 10-20% MeOH/CH₂Cl₂ with 0-10% NH₄OH added toyield compound 9 as a white solid (0.098 g). ¹H NMR δ (DMSO, 400 MHz),11.22 (s, 1H), 7.79 (s, 1H), 7.44 (d, J=8.5 Hz, 1H), 7.29 (m, 2H), 4.70(s, 1H), 3.92 (m, 1H), 3.66 (m, 2H), 3.34 (m, 6H), 2.90 (m, 1H), 2.75(m, 1H), 2.60 (m, 1H), 2.40 (m, 1H), 2.25 (m, 2H), 2.20 (m, 3H), 1.84(m, 1H), 1.72 (m, 3H), 1.41 (m, 7H), 1.07 (d, J=6.6 Hz, 3H). MS (ES):522 [M+H].

Example 7

[0216]

[0217] To a mixture of acid 7 (0.075 g, 0.16 mmol), DMF (2 drops) inCH₂Cl₂ (2 mL) was added (COCl)₂ (0.8 mL, 2 M/CH₂Cl₂, 1.6 mmol). When thegas release ceased, the mixture was placed under high vacuum to obtain asolid. To this solid was added CH₂Cl₂ (2 mL), (S)(+)2-(aminomethyl)-pyrrolidine (0.20 mL, 1.87 mmol) and NEt₃ (0.170 mL, 1.2mmol). The mixture was stirred for 1 h at r.t., poured into sat. NaHCO₃and extracted with EtOAc. The organic layer was separated, washed withbrine, dried with anhydrous Na₂SO₄, concentrated by rotary evaporationand purified by flash chromatography on silica gel with a gradientelution of 20-40% MeOH/CH₂Cl₂ in 0-10% NH₄OH to yield compound 10 as ayellowish solid (0.053 g). ¹H NMR δ (DMSO, 400 MHz), 11.21 (s, 1H), 7.78(s, 1H), 7.58 (m, 1H), 7.44 (d, J=8.5 Hz, 1H), 7.28 (d, J=8.5 Hz, 1H),3.67 (m, 2H), 3.1-3.5 (m, 8H), 2.75-2.9 (m, 4H), 2.6 (m, 1H), 2.38 (m,1H), 2.22 (m, 2H), 2.01 (m, 2H), 1.6-1.9 (m, 8H), 1.4 (m, 7H), 1.47 (m,1H). MS (ES): 547 [M+H].

Example 8

[0218]

[0219] A mixture of amide 8 (0.695 g, 1.50 mmol) and POCl₃ (0.42 mL, 4.5mmol) in anhydrous pyridine (14 mL) was heated to 120° C. in a sealedvessel for 2 h. It was cooled to r.t., poured into saturated NaHCO₃solution and extracted with EtOAc. The organic layer was separated,washed with brine, dried with anhydrous Na₂SO₄, concentrated by rotaryevaporation and purified by flash chromatography on silica gel with agradient elution of 5-30% MeOH/CH₂Cl₂ to yield the corresponding nitrileas a yellowish solid (0.400 g).

[0220] The above nitrile was heated with Bu₃SnN₃ (0.74 mL, 2.7 mmol) intoluene (3 mL) at 120° C. in a sealed vessel for 2 days. At thecompletion of the reaction, the reaction mixture was cooled to r.t.,acidified with 1M HCl in ether and purified by flash chromatography onsilica gel with a gradient elution of 20-40% MeOH/CH₂Cl₂ in 0-10% NH₄OHto yield compound 11 as a yellowish solid (0.196 g). ¹H NMR δ (DMSO, 400MHz), 11.26 (s, 1H), 7.79 (s, 1H), 7.44 (d, J=8.5 Hz, 1H), 7.30 (d,J=8.5 Hz, 1H), 3.37 (m, 3H), 3.14 (m, 4H), 3.14 (m, 1H), 2.77 (m, 1H),2.64 (m, 3H), 2.43 (m, 2H), 2.25 (m, 3H), 2.01 (m, 2H), 1.91 (m, 1H),1.76 (m, 2H), 1.55 (m, 1H), 1.35-1.5 (m, 4H). MS (ES): 489 [M+H].

Example 9

[0221]

[0222] Compound 12 was prepared following the procedures detailed inExample 4, substituting 4-(2-oxo-ethyl)-tetrahydro-pyran-4-carboxylicacid methyl ester with cyclohexane carboxylic acid methyl ester. ¹H NMRδ (d₆-DMSO) 12.47 (bs, 1H), 11.31 (s, 1H), 10.46 (bs, 1H), 7.78 (s, 1H),7.44 (d, J=8.5 Hz, 1H), 7.28 (dd, J=8.5, 1.4 Hz, 1H), 3.78 (d, J=11.6Hz, 1H), 3.56 (d, J=11.6 Hz, 1H), 3.10 (td, J=12.2, 5.0 Hz, 1H), 2.94(m, 2H), 2.81 (d, J=14.4 Hz, 2H), 2.65 (t, J=5.7 Hz, 1H), 2.40 (dd,J=15.7, 9.8 Hz, 1H), 2.05-1.90 (m, 5H), 1.80-1.65 (m, 3H), 1.54-1.39 (m,3H), 1.39 (d, J=6.6 Hz, 3H), 1.38-1.22 (m, 5H). MS (ES): 463 [M+H].

Example 10

[0223]

[0224] Compound 13 was synthesized in 5 steps according to the followingscheme.

[0225] Step 1. A mixture of 4-allyl-tetrahydropyran carboxylic acidmethyl ester (15.7 g, 92.4 mmol) and LiOH.H₂O (29 g, 688 mmol) in THF(110 mL), MeOH (110 mL) and water (5 mL) was heated to 85° C. in asealed vessel overnight. Upon cooling to r.t., it was extracted withEtOAc, washed with water, dried with anhydrous Na₂SO₄ and concentratedby rotary evaporation to yield the corresponding carboxylic acid as awhite solid (12.82 g).

[0226] Step 2. To a mixture of the above acid (12.82 g, 75.4 mmol), DMF(4 drops) in CH₂Cl₂ (300 mL) was added (COCl)₂ (75.4 mL, 2 M in CH₂Cl₂,150.8 mmol). When the gas release ceased, the mixture was placed on arotary evaporator. The obtained acid chloride (in 150 mL of dry acetone)was added to NaN₃ (48.75 g, 0.75 mol, in 300 mL of water) at 0° C. over30 min. After stirring 2 h at r.t., the mixture was poured intoice-water and extracted with ether. The organic layer was separated,washed with brine, dried over anhydrous Na₂SO₄ and concentrated byrotary evaporation. The obtained azide was dissolved in 100 mL ofbenzene and added slowly to 100 mL of refluxing benzene. The refluxcontinued for another 40 min at which time no more gas was released.Benzene was distilled off and 120 mL of MeOH was added to the mixture.The mixture was heated at refluxing for 36 h, cooled to r.t. and wasdirectly chromatographed using 40% EtOAc/hexanes as eluent. Thecarbamate product was obtained as a white solid (14.15 g).

[0227] Step 3. The obtained carbamate (0.498 g, 2.5 mmol) was stirredwith NaIO₄ (1.18 g, 5.5 mmol) and OsO₄ (30 mg) in MeOH (5 mL) and H₂O (5mL) for 15 min. The mixture was directly loaded onto a column forchromatography with a gradient elution of 70-90% EtOAc/hexanes as theeluent to yield the aldehyde product as an oil (0.48 g).

[0228] Step 4. The above aldehyde (0.38 g, 1.9 mmol) was stirred withamine 4 (0.587 g, 1.9 mmol) and NaBH(OAc)₃ (1.61 g, 7.6 mmol) inClCH₂CH₂Cl (15 mL) overnight. The reaction mixture was poured into adilute aqueous ammonia solution and extracted with EtOAc. The organiclayer was separated, washed with brine, dried with anhydrous Na₂SO₄,concentrated by rotary evaporation and purified by flash chromatographyon silica gel with a gradient elution of 20-40% MeOH/CH₂Cl₂ with 0-10%NH₄OH added to yield the coupled product as a solid (0.618 g).

[0229] Step 5. A mixture of the obtained product (0.618 g, 1.25 mmol)and LiOH.H₂O (3.0 g, 71.5 mmol) in dioxane (20 mL) and water (10 mL) washeated to 120° C. in a sealed vessel for 8 h. Upon cooling to r.t., itwas poured into saturated NaHCO₃ and extracted with EtOAc. The organiclayer was separated, washed with brine, dried with anhydrous Na₂SO₄,concentrated by rotary evaporation and purified by flash chromatographyon silica gel with a gradient elution of 20-40% MeOH/CH₂Cl₂ with 0-10%NH₄OH added to yield compound 13 as a yellowish solid (0.268 g). MS(ES): 436 [M+H].

Example 11

[0230]

[0231] To a mixture of amine 13 (0.096 g, 0.22 mmol) and NEt₃ (0.084 mL,0.6 mmol) in CH₂Cl₂ (1.5 mL) was added trifluoromethanesulfonylanhydride (0.067 mL, 0.40 mmol) at 0 C. After 20 min, it was poured intosaturated NaHCO₃ solution and extracted with EtOAc. The organic layerwas separated, washed with brine, dried with anhydrous Na₂SO₄,concentrated by rotary evaporation and purified by flash chromatographyon silica gel with a gradient elution of 5-20% MeOH/CH₂Cl₂ to yieldcompound 14 as a yellowish solid (0.077 g). ¹H NMR δ (DMSO, 400 MHz),11.27 (s, 1H), 7.81 (s, 1H), 7.46 (d, J=8.5 Hz, 1H), 7.30 (d, J=8.5 Hz,1H), 3.5-4.1 (m, 10H), 2.95 (m, 2H), 2.83 (m, 1H), 2.80 (m, 1H), 2.69(m, 1H), 2.35 (m, 2H), 2.00 (m, 5H), 1.73 (m, 3H), 1.40 (d, J=6.5 Hz,3H). MS (ES): 568 [M+H].

Example 12

[0232]

[0233] To a mixture of acid 7 (0.070 g, 0.15 mmol), DMF (2 drops) andDCM (2 mL) was added (COCl)₂ (0.6 mL, 2 M/DCM, 1.2 mmol). When the gasrelease ceased, solvents were evaporated to obtain a solid. To thissolid was added DCM (2 mL), morpholine (0.2 mL, 2.3 mmol) and TEA (0.15mL, 1.1 mmol). The mixture was stirred at r.t. for 1 h, poured intosaturated NaHCO₃ and extracted with EtOAc. The organic layer wasseparated, washed with brine, dried with anhydrous Na₂SO₄, concentratedby rotary evaporation and purified by flash chromatography on silica geleluted with 20% MeOH/DCM to yield 15 as a yellowish solid (0.080 g). ¹HNMR δ (DMSO, 500 MHz): 11.20 (s, 1H), 7.79 (s, 1H), 7.44 (d, J=8.5 Hz,1H), 7.28 (d, J=8.5 Hz, 1H), 3.69 (m, 2 H), 3.58 (m, 8H), 3.44 (m, 2 H),3.32 (m, 2H), 3.24 (m, 1H), 2.90 (m, 1H), 2.74 (m, 1H), 2.42 (m, 1H),2.23 (m, 2H), 2.08 (m, 2H), 1.86 (m, 4H), 1.68 (m, 1H), 1.52 (m, 2H),1.29 (m, 2H), 1.27 (d, J=8.5 Hz, 3H). MS (ES): 534 [M+H].

Example 13

[0234]

[0235] A mixture of acid 7 (0.306 g, 0.66 mmol),4-(2-aminoethyl)morpholine (0.375 mL, 2.64 mmol), EDC.HCl (0.381 g, 1.98mmol), HOBt (0.267 g, 1.98 mmol), NMP (0.44 mL, 4 mmol), DCM (5 mL) andDMF (5 mL) was stirred at r.t. for 3 h. The mixture was poured intosaturated NaHCO₃ and extracted with EtOAc. The organic layer wasseparated, washed with brine, dried with anhydrous Na₂SO₄, concentratedby rotary evaporation and purified by flash chromatography on silica gelwith a gradient elution of 10-20% MeOH/DCM containing 1-3% NH₄OH toyield 16 as a white solid (0.310 g). MS (ES): 577 [M+H].

Example 14

[0236]

[0237] A mixture of acid 7 (0.102 g, 0.22 mmol),2-aminoethyl-1-ethylpyrrolidine (0.128 g, 1 mmol), EDC.HCl (0.127 g,0.66 mmol), HOBt (0.089 g, 0.66 mmol), NMP (0.22 mL, 2 mmol), DCM (1.5mL) and DMF (1.5 mL) was stirred at r.t. for 8 h. The mixture was pouredinto saturated NaHCO₃ and extracted with EtOAc. The organic layer wasseparated, washed with brine, dried with anhydrous Na₂SO₄, concentratedby rotary evaporation and purified by flash chromatography on silica gelwith a gradient elution of 10-25% MeOH/DCM containing 1-3% NH₄OH toyield 17 as a white solid (0.095 g). ¹H NMR δ (DMSO, 500 MHz): 11.19 (s,1H), 7.78 (s, 1H), 7.60 (t, J=5.5 Hz, 1H), 7.43 (d, J=8.5 Hz, 1H), 7.28(d, J=8.5 Hz, 1H), 3.66 (m, 2H), 3.30 (m, 1H), 3.21 (m, 1H), 3.02 (m,1H), 2.60-3.0 (m, 3H), 2.73 (m, 1H), 2.60 (m, 1H), 2.47 (m, 1H), 2.40(m, 1H), 2.20 (m, 3H), 2.10 (m, 1H), 2.00 (m, 2H), 1.79-1.95 (m, 4H),1.5-1.7 (m, 6H), 1.40 (m, 5H), 1.37 (d, J=6.5 Hz, 3H), 1.06 (m, 1H),1.04 (t, J=7.5 Hz, 3H). MS (ES): 575 [M+H].

Example 15

[0238]

[0239] To a mixture of acid 7 (0.325 g, 0.7 mmol), DMF (2 drops) and DCM(8 mL) was added (COCl)₂ (3.6 mL, 2 M in DCM, 7.2 mmol). When the gasrelease ceased, the mixture was placed under high vacuum to obtain asolid. To this solid was added DCM (8 mL), MeONH₂.HCl (0.800 g, 9.6mmol) and TEA (0.60 mL, 4.3 mmol). The mixture was stirred at r.t. for 1h, poured into saturated NaHCO₃ and extracted with EtOAc. The organiclayer was separated, washed with brine, dried with anhydrous Na₂SO₄,concentrated by rotary evaporation and purified by flash chromatographyon silica gel eluted with a gradient elution of 20-40% MeOH/DCM mixedwith 0-10% NH₄OH to yield 18 as an off-white solid (0.24 g). ¹H NMR δ(DMSO, 500 MHz): 11.20 (s, 1H), 11.15 (s, 1H), 7.79 (s, 1H), 7.43 (d,J=8.5 Hz, 1H), 7.28 (d, J=8.5 Hz, 1H), 3.68 (m, 2H), 3.63 (s, 3H), 3.35(m, 2H), 3.23 (m, 1H), 2.89 (m, 1H), 2.75 (m, 1H), 2.60 (m, 1H), 2.40(m, 1H), 2.26 (m, 2H), 1.87-2.0 (m, 3H), 1.81 (m, 1H), 1.69 (m, 3H),1.43 (m, 4H), 1.38 (d, J=6.5 Hz, 3H), 1.19 (m, 1H). MS (ES): 494 [M+H].

Example 16

[0240]

[0241] To a mixture of acid 7 (0.060 g, 0.13 mmol), DMF (1 drop) and DCM(2 mL) was added (COCl)₂ (0.6 mL, 2 M in DCM, 1.2 mmol). When the gasrelease ceased, the mixture was placed under high vacuum to obtain asolid. To this solid was added DCM (2 mL), NH₂CH₂CN (0.200 g, 3.6 mmol)and TEA (0.15 mL, 1.07 mmol). After stirring at r.t. for 1 h, themixture was poured into saturated NaHCO₃ and extracted with EtOAc. Theorganic layer was separated, washed with brine, dried with anhydrousNa₂SO₄, concentrated by rotary evaporation and purified by flashchromatography on silica gel eluted with a gradient elution of 20-40%MeOH/DCM mixed with 0-8% NH₄OH to yield 19 as a white solid (0.028 g).¹H NMR δ (DMSO, 500 MHz): 11.20 (s, 1H), 8.50 (s, 1H), 7.78 (s, 1H),7.43 (d, J=8.5 Hz, 1H), 7.28 (d, J=8.5 Hz, 1H), 4.15 (d, J=5.5 Hz, 2H),3.69 (m, 2H), 3.33 (s, 3H), 3.23 (m, 1), 2.88 (m, 1H), 2.75 (m, 1H),2.60 (m, 1H), 2.40 (m, 1H), 2.22 (m, 2H), 2.00 (m, 2H), 1.6-1.95 (m,4H), 1.50 (m, 2H), 1.39 (m, 2H), 1.37 (d, J=6.5 Hz, 3H), 1.28 (m, 1H).MS (ES): 503 [M+H].

Example 17

[0242]

[0243] A mixture of acid 7 (0.102 g, 0.22 mmol),2-aminomethyl-2-propanol (0.095 g, 0.88 mmol, prepared according to Rai,B.; Dekhordi, L. S.; Khodr, H.; Jin, Y.; Liu, Z.; R. C. Hider (1998) J.Med. Chem. 41:3347-3359), EDC-HCl (0.127 g, 0.66 mmol), HOBt (0.089 g,0.66 mmol), NMP (0.11 mL, 1 mmol), DCM (2 mL) and DMF (2 mL) was stirredat r.t. overnight. The mixture was poured into saturated NaHCO₃ andextracted with EtOAc. The organic layer was separated, washed withbrine, dried with anhydrous Na₂SO₄, concentrated by rotary evaporationand purified by flash chromatography on silica gel with a gradientelution of 20-40% MeOH/DCM mixed with 0-10% NH₄OH to yield 20 as a whitesolid (0.085 g). ¹H NMR δ (DMSO, 500 MHz): 11.20 (s, 1 H), 7.79 (s, 1H), 7.54 (m, 1 H), 7.44 (d, J=8.5 Hz, 1H), 7.28 (d, J=8.5 Hz, 1H), 4.51(s, 1H), 3.68 (m, 2H), 3.38 (m, 2H), 3.26 (m, 1H), 3.13 (d, J=6.0 Hz,1H), 2.90 (m, 1H), 2.75 (m, 1H), 2.60 (m, 1H), 2.40 (m, 1H), 2.28 (m,2H), 2.00 (m, 2H), 1.80 (m, 5H), 1.45 (m, 4H), 1.40 (d, J=6.5 Hz, 3H),1.29 (m, 1H), 1.09 (s, 6H). MS (ES): 536 [M+H].

Example 18

[0244]

[0245] Synthesized in two steps: To a mixture of acid 7 (1.00 g, 2.2mmol), DMF (3 drops) and DCM (20 mL) was added (COCl)₂ (10 mL, 2 M inDCM, 20 mmol). When the gas release ceased, the mixture was placed undera rotary evaporator followed by a high vacuum pump to obtain thecorresponding acyl chloride as a solid. The acyl chloride (dissolved in10 mL of DCM and 5 mL of DMF) was added to a flask containingNH₂CH₂CH₂NH₂ (4.42 mL, 66 mmol) in DCM (15 mL). The mixture was stirredat r.t. for 1 h, poured into saturated NaHCO₃ and extracted with EtOAc.The organic layer was separated, washed with brine, dried with anhydrousNa₂SO₄, concentrated by rotary evaporation and purified by flashchromatography on silica gel eluted with a gradient elution of 20-40%MeOH/DCM mixed with 0-10% NH₄OH to yield the corresponding aminoethylamide as a yellowish solid.

[0246] The above amide (0.035 g, 0.07 mmol) was stirred with AcCl (0.01mL, 0.14 mmol) and TEA (0.035 mL, 0.25 mmol) in DCM (1 mL) for 10 min.The mixture was directly loaded onto a silica gel column eluted with agradient elution of 20-40% MeOH/DCM mixed with 0-7% NH₄OH to yield 21 asa yellowish solid (0.022 g). MS (ES): 549 [M+H].

Example 19

[0247]

[0248] Synthesized in two steps: A mixture of the intermediateaminoethyl amide of Example 18 (0.67 g, 1.65 mmol), trifluoroaceticanhydride (0.292 mL, 2.1 mmol), TEA (0.42 mL, 3 mmol) and DCM (10 mL)was stirred at r.t. for 10 min. The mixture was poured into saturatedNaHCO₃ and extracted with EtOAc. The organic layer was separated, washedwith brine, dried with anhydrous Na₂SO₄, concentrated by rotaryevaporation and purified by flash chromatography on silica gel with agradient elution of 10-20% MeOH/DCM mixed with 1-2% NH₄OH to yield thecorresponding trifluoromethyl acetamide as a yellowish solid (0.510 g).

[0249] The above amide (0.300 g, 0.5 mmol) was refluxed with LAH (0.050g, 1.3 mmol) in THF (5 mL) for 1 h. At this time HPLC-MS indicated halfcompletion of the reaction. A second portion of LAH (0.050 g, 1.3 mmol)was added. After refluxing for another 1 h, no more progress wasobserved. The mixture was poured into dilute ammonium hydroxide andextracted with EtOAc. The organic layer was separated, washed withbrine, dried with anhydrous Na₂SO₄, concentrated by rotary evaporation.Flash chromatography on silica gel with a gradient elution of 20-40%MeOH/DCM mixed with 1-10% NH₄OH afforded no separation. Finallyseparation was achieved by preparative HPLC to yield 22 as a white solid(0.037 g). MS (ES): 589 [M+H].

Example 20

[0250]

[0251] A mixture of amine 13 (0.090 g, 0.2 mmol), tetrahydro-3-furoicacid (0.047 g, 0.4 mmol), EDC.HCl (0.115 g, 0.6 mmol), HOBt (0.081 g,0.6 mmol), TEA (0.140 mL, 1 mmol), DCM (1 mL) and DMF (1 mL) was stirredat r.t. overnight. The mixture was poured into saturated NaHCO₃ andextracted with EtOAc. The organic layer was separated, washed withbrine, dried with anhydrous Na₂SO₄, concentrated by rotary evaporationand purified by flash chromatography on silica gel with a gradientelution of 20-40% MeOH/DCM mixed with 0-10% NH₄0H to yield 23 as ayellowish solid (0.069 g). ¹H NMR δ (DMSO, 500 MHz): 11.20 (s, 1H), 7.78(s, 1H), 7.47 (s, br, 1H), 7.43 (d, J=8.5 Hz, 1H), 7.28 (d, J=8.5 Hz,1H), 3.89 (t, J=7.8 Hz, 1H), 3.58-3.85 (m, 6H), 3.48 (m, 2H), 3.30 (m,1H), 3.05 (m, 1H), 2.74 (m, 1H), 2.62 (m, 1H), 2.51 (m, 1H), 2.35 (m,1H), 2.09 (m, 2H), 2.00 (m, 2H), 1.92 (m, 2H), 1.84 (m, 1H), 1.4-1.55(m, 3H), 1.37 (d, J=6.5 Hz, 3H), 1.30 (m, 1H). MS (ES): 534 [M+H].

Example 21

[0252]

[0253] A mixture of amine 13 (0.065 g, 0.15 mmol), methanesulfonylacetic acid (0.061 g, 0.45 mmol), EDC.HCl (0.086 g, 0.45 mmol), HOBt(0.061 g, 0.45 mmol), NMP (0.165 mL, 1.5 mmol), DCM (1.5 mL) and DMF(1.5 mL) was stirred at r.t. for 24 h. The mixture was poured intosaturated NaHCO₃ and extracted with EtOAc. The organic layer wasseparated, washed with brine, dried with anhydrous Na₂SO₄, concentratedby rotary evaporation and purified by flash chromatography on silica gelwith a gradient elution of 15-20% MeOH/DCM mixed with 1-2% NH₄OH toyield 24 as a white solid (0.052 g). ¹H NMR δ (DMSO, 500 MHz): 11.19 (s,1H), 7.90 (s, 1H), 7.79 (s, 1H), 7.44 (d, J=8.5 Hz, 1H), 7.28 (d, J=8.5Hz, 1H), 4.13 (s, 2H), 3.63 (m, 2H), 3.54 (m, 2H), 3.32 (m, 2H), 3.11(s, 3H), 2.92 (m, 1H), 2.74 (m, 1H), 2.60 (m, 1H), 2.40 (m, 3H), 2.08(m, 2H), 1.92 (m, 2H), 1.83 (m, 1H), 1.69 (m, 1H), 1.54 (m, 2H), 1.40(m, 2H), 1.38 (d, J=6.5 Hz, 3H), 1.19 (m, 1H). MS (ES): 556 [M+H].

Example 22

[0254]

[0255] A mixture of amine 13 (0.097 g, 0.224 mmol),4,4-dioxo-tetrahydrothiopyranyl carboxylic acid (0.040 g, 0.224 mmol,prepared as following), EDC.HCl (0.107 g, 0.56 mmol), HOBt (0.076 g,0.56 mmol), NMP (0.275 mL, 2.5 mmol), DCM (1.5 mL) and DMF (1.5 mL) wasstirred at r.t. overnight. The mixture was poured into saturated NaHCO₃and extracted with EtOAc. The organic layer was separated, washed withbrine, dried with anhydrous Na₂SO₄, concentrated by rotary evaporationand purified by flash chromatography on silica gel with a gradientelution of 20% MeOH/DCM mixed with 1-3% NH₄0H to yield 25 as a whitesolid (0.074 g). ¹H NMR δ (DMSO, 500 MHz): 11.19 (s, 1H), 7.78 (s, 1H),7.50 (s, 1H), 7.43 (d, J=8.5 Hz, 1H), 7.28 (d, J=8.5 Hz, 1H), 3.62 (m,2H), 3.48 (m, 2H), 3.25 (m, 1H), 3.14 (m, 4H), 2.90 (m, 1H), 2.74 (m,1H), 2.58 (m, 3H), 2.2-2.45 (m, 3H), 2.09 (m, 6H), 1.8-1.98 (m, 4H),1.65 (m, 1H), 1.43 (m, 3H), 1.38 (d, J=6.5 Hz, 3H), 1.28 (m, 1H). MS(ES): 596 [M+H].

[0256] Preparation of 4,4-dioxo-tetrahydrothiopyranyl carboxylic acid: Amixture of 2,2-dimethyl-1,3-dioxane-4,6-dione (10 g, 69.4 mmol), vinylsulfone (8.19 g, 69.4 mmol), KOH (9.72 g, 173.6 mmol) and tBuOH (140 mL)was refluxed overnight. The supernant was decanted and 120 mL of 20%aqueous H₂SO₄ was added to the residual solid. The obtained mixture wasrefluxed for another 3 h, extracted with iPrOH/CHCl₃, dried withanhydrous Na₂SO₄ and concentrated by rotary evaporation to afford thedesired 4,4-dioxo-tetrahydrothiopyranyl carboxylic acid.

Example 23

[0257]

[0258] Synthesized in two steps: A mixture of amine 13 (0.100 g, 0.23mmol), ClSO₂CH₂CO₂Me (0.039 g, 0.3 mmol), pyrindine (0.49 mL, 0.6 mmol)and DCM (2 mL) was stirred at r.t. for 20 min. The mixture was pouredinto saturated NaHCO₃ and extracted with EtOAc. The organic layer wasseparated, washed with brine, dried with anhydrous Na₂SO₄, concentratedby rotary evaporation and purified by prep. HPLC to yield thecorresponding sulfonamide.

[0259] The above sulfonamide (0.020 g, 0.038 mmol) was reacted with LAH(0.20 mL, 1 M in THF, 0.2 mmol) in THF (1 mL) at r.t. for 15 min. Thereaction mixture was poured into dilute ammonium hydroxide and extractedwith EtOAc. The organic layer was separated, washed with brine, driedwith anhydrous Na₂SO₄, concentrated by rotary evaporation. Flashchromatography on silica gel with a gradient elution of 15-20% MeOH/DCMmixed with 1-3% NH₄OH afforded compound 26 as a white solid (0.004 g).MS (ES): 544 [M+H].

Example 24

[0260]

[0261] A mixture of amine 13 (0.239 g, 0.55 mmol), C(Me)₂(OH)CF₂CO₂H(0.085 g, 0.55 mmol, prepared according to Dolbier, Jr. W. R.; Ocampo,R. (1995) J. Organic Chem. 60:5378 and Hallinan, E. A.; Fried, J. (1984)Tetrahedron Lett. 25:2301), EDC-HCl (0.264 g, 1.37 mmol), HOBt (0.186 g,1.37 mmol), NMP (0.44 mL, 4 mmol), DCM (3 mL) and DMF (3 mL) was stirredat r.t. for 24 h. The mixture was poured into saturated NaHCO₃ andextracted with EtOAc. The organic layer was separated, washed withbrine, dried with anhydrous Na₂SO₄, concentrated by rotary evaporationand purified by flash chromatography on silica gel with a gradientelution of 15-20% MeOH/DCM mixed with 0-2% NH₄OH to yield 27 as a whitesolid (0.098 g). ¹H NMR δ (DMSO, 500 MHz): 11.19 (s, 1H), 7.79 (m, 2H),7.44 (d, J=8.5 Hz, 1H), 7.28 (d, J=8.5 Hz, 1H), 5.59 (s, 1H), 3.63 (m,2H), 3.52 (m, 2H), 3.25 (m, 1H), 2.93 (m, 1H), 2.75 (m, 1H), 2.61 (m,1H), 2.40 (m, 3H), 2.25 (m, 2H), 1.94 (m, 3H), 1.81 (m, 1H), 1.66 (m,1H), 1.56 (m, 2H), 1.42 (m, 2H), 1.38 (d, J=6.5 Hz, 3H), 1.19 (m, 7H).MS (ES): 572 [M+H].

Example 25

[0262]

[0263] Step 1. A mixture of amine 13 (0.654 g, 1.5 mmol),OHCCH₂CH₂CH₂CO₂Et (0.260 g, 2 mmol), NaBH(OAc)₃ (1.27 g, 6 mmol) andClCH₂CH₂Cl (10 mL) was stirred at r.t. overnight. The mixture was pouredinto dilute NH₄OH and extracted with EtOAc. The organic layer wasseparated, washed with brine, dried with anhydrous Na₂SO₄, concentratedby rotary evaporation and purified by flash chromatography on silica gelwith a gradient elution of 15-20% MeOH/DCM mixed with 1-4% NH₄OH toyield the corresponding monoalkylated amine (0.35 g).

[0264] The product obtained above (0.35 g, 0.63 mmol) was hydrolyzed bytreating with LiOH.H₂O (0.50 g, 12 mmol) in dioxane (4 mL) and H₂O (2mL) at r.t. for 3 h. The mixture was acidified with HOAc to slightlyacidic, and was concentrated to dryness.

[0265] The above acid (˜0.1 mmol, contained inorganic salt) was heatedwith NaOAc (0.200 g) in Ac₂O (2 mL) at 105° C. for 20 min. The reactionmixture was poured into saturated NaHCO₃ and extracted with EtOAc. Theorganic layer was separated, washed with brine, dried with anhydrousNa₂SO₄, concentrated by rotary evaporation and purified by flashchromatography on silica gel with a gradient elution of 15-20% MeOH/DCMmixed with 0-3% NH₄OH to yield 28 as a white solid (0.040 g). ¹H NMR δ(DMSO, 500 MHz): 11.19 (s, 1H), 7.78 (s, 1H), 7.43 (d, J=8.5 Hz, 1H),7.28 (d, J=8.5 Hz, 1H), 3.64 (m, 2H), 3.45 (m, 4H), 3.36 (m, 1H), 2.92(m, 1H), 2.75 (m, 1H), 2.61 (m, 1H), 2.38 (m, 5H), 2.26 (t, J=8.0 Hz,2H), 1.89 (m, 6H), 1.68 (m, 1H), 1.41 (m, 2H), 1.38 (d, J=6.5 Hz, 3H),1.26 (m, 1H). MS (ES): 504 [M+H].

Example 26

[0266]

[0267] Synthesized in two steps: A mixture of amine 13 (0.664 g, 1.52mmol), 2,5-dimethoxy-3-tetrahydrofuran-carboxaldehyde (0.487 g, 3.0mmol) and HOAc (8 mL) was heated to 70° C. for 2 h. The mixture wascooled to r.t., basified with saturated NaHCO₃ and extracted with EtOAc.The organic layer was separated, washed with brine, dried with anhydrousNa₂SO₄, concentrated by rotary evaporation and purified by flashchromatography on silica gel with a gradient elution of 10-20% MeOH/DCMmixed with 0-2% NH₄OH to yield the corresponding formylpyrrolederivative (0.70 g).

[0268] The above aldehyde was converted to nitrile by the followingreaction. In a vial containing NH₂OH.HCl (0.083 g, 1.2 mmol) and CH₃CN(3 mL) at 0° C. was added TEA (0.168 mL, 1.2 mmol) and the aldehyde(0.470 g, 0.91 mmol). The mixture was stirred at 0° C. for 30 min. andat r.t. for 4 h. At this time, phthalic anhydride (0.178 g, 1.2 mmol)was added and the mixture was heated to 90° C. for 1 h, poured intosaturated NaHCO₃ and extracted with EtOAc. The organic layer wasseparated, washed with brine, dried with anhydrous Na₂SO₄, concentratedby rotary evaporation and purified by flash chromatography on silica gelwith a gradient elution of 10-20% MeOH/DCM mixed with 0-1% NH₄OH toyield 29 as a yellowish solid (0.167 g). ¹H NMR δ (DMSO, 500 MHz): 11.18(s, 1H), 7.84 (s, 1H), 7.77 (s, 1H), 7.43 (d, J=8.5 Hz, 1H), 7.28 (d,J=8.5 Hz, 1H), 7.17 (m, 1H), 6.54 (m, 1H), 3.71 (m, 2H), 3.41 (m, 2H),3.08 (m, 1H), 2.73 (m, 2H), 2.55 (m, 2H), 2.38 (m, 1H), 2.25 (m, 2H),1.85-2.1 (m, 6H), 1.77 (m, 2H), 1.54 (m, 1H), 1.36 (d, J=6.5 Hz, 3H),1.23 (m, 2H). MS (ES): 511 [M+H].

Example 27

[0269]

[0270] Synthesized in three steps: A mixture of amine 13 (0.600 g, 1.38mmol), BocNHCH₂CHO (0.220 g, 0.38 mmol), NaBH(OAc)₃ (1.17 g, 5.5 mmol)and ClCH₂CH₂Cl (14 mL) was stirred at r.t. overnight. The mixture waspoured into dilute NH₄OH and extracted with EtOAc. The organic layer wasseparated, washed with brine, dried with anhydrous Na₂SO₄, concentratedby rotary evaporation and purified by flash chromatography on silica gelwith a gradient elution of 15-20% MeOH/DCM mixed with 1-4% NH₄OH toyield the corresponding reductive amination product (0.311 g).

[0271] The product obtained above (0.311 g, 0.537 mmol) was stirred withHCl (1.5 mL, 4 M in dioxane, 6 mmol) in 1.5 mL of DCM for 30 min. Themixture was poured into dilute NH₄OH and extracted with EtOAc. Theorganic layer was separated, washed with brine, dried with anhydrousNa₂SO₄, concentrated by rotary evaporation to yield the de-protectedproduct which was directed used for the next step.

[0272] The resulted de-Boc product (0.129 g, 0.27 mmol) was refluxedwith carbonyldiimidazole (0.087 g, 0.54 mmol) in DCM (8 mL) for 1 h. Thereaction mixture was poured into saturated NaHCO₃ and extracted withEtOAc. The organic layer was separated, washed with brine, dried withanhydrous Na₂SO₄, concentrated by rotary evaporation and purified byflash chromatography on silica gel with a gradient elution of 15-20%MeOH/DCM mixed with 1-2% NH₄OH to yield 30 as a white solid (0.123 g).MS (ES): 505 [M+H].

Example 28

[0273]

[0274] A sample of the alcohol compound from Example 3 was converted tothe corresponding aldehyde by oxidation with SO₃.Py and DMSO. To thesolution of this aldehyde (0.100 g, 0.2 mmol) in MeOH (1 mL) at 0° C.was added glyoxal (0.055 g, 0.95 mmol, 50% in water) followed by NH₃(0.50 mL, 1 mmol, 2 M in MeOH). The mixture was allowed to warm to r.t.and stirred for 24 h. The mixture was poured into with saturated NaHCO₃and extracted with EtOAc. The organic layer was separated, washed withbrine, dried with anhydrous Na₂SO₄, concentrated by rotary evaporationand purified by flash chromatography on silica gel with a gradientelution of 15-20% MeOH/DCM with 1-4% NH₄OH added to yield 31 as ayellowish solid (0.027 g). ¹H NMR δ (DMSO, 500 MHz): 11.60 (s, br, 1H),11.20 (s, 1H), 7.77 (s, 1H), 7.43 (d, J=8.5 Hz, 1H), 7.28 (d, J=8.5 Hz,1H), 6.96 (s, br, 2H), 3.70 (m, 2H), 3.32 (m, 4H), 3.19 (m, 1H), 2.88(s, 1H), 2.72 (m, 1H), 2.57 (m, 1H), 2.51 (m, 1H), 2.38 (m, 1H), 2.23(m, 2H), 2.09 (m, 1H), 1.82 (m, 3H), 1.66 (m, 2H), 1.39 (m, 2H), 1.65(d, J=6.5 Hz, 3H), 1.19 (m, 1H). MS (ES): 487 [M+H].

Example 29

[0275]

[0276] Synthesized in two steps: To a solution of diazomethylphosphonate(0.180 g, 1.2 mmol, Seyferth, D.; Marmor, R. S.; Hilbert, P. (1971) J.Organic Chem. 36:1379) in THF (6 mL) cooled to −78° C. under a nitrogenatmosphere was added KOtBu (1.8 mL, 1.8 mmol, 1.0 M in THF) dropwise.The obtained mixture was stirred for 10 min. at the low temperature. Analdehyde (0.268 g, 0.6 mmol, dissolved in THF, same one used as for thepreparation of compound 31) was added to the above mixture dropwise. Themixture was stirred for 30 min. at −78° C. and another 30 min. at r.t.The reaction was quenched with water, and the mixture was poured intosaturated NaHCO₃ and extracted with EtOAc. The organic layer wasseparated, washed with brine, dried with anhydrous Na₂SO₄, concentratedby rotary evaporation and purified by flash chromatography on silica geleluted with gradient elution of 5-20% MeOH/DCM to yield thecorresponding alkyne as a white solid (0.200 g).

[0277] The obtained alkyne (0.100 g, 0.22 mmol) was heated with TMSN₃ (1mL) in a sealed vial at 140° C. for two days. The whole mixture wasdirectly loaded onto a column, eluted with 15-20% MeOH/DCM mixed with1-5% NH₄OH to yield 32 as a white solid (0.0048 g). MS (ES): 488 [M+H].

Example 30

[0278]

[0279] Lithium diisopropylamide (1.1 mL,2.0 M, 2.2 mmol) was added to asolution of THF (0.25 M solution) containingN-allyl-N-t-butoxycarbonyl-methanesulfonamide (0.518 g, 2.2 mmol) at−78° C. After stirring for 50 min. at −78° C., the solution of thealdehyde intermediate from Example 28 in dry THF (0.470 g, 1.05 mmol)was added and the mixture was stirred at −78° C. to room temperatureovernight. The reaction was treated with saturated sodium bicarbonatesolution and extracted with ethyl acetate, the organic layer was washedwith brine and dried, concentrated and purified by flash chromatographyon silica gel eluted with 3% MeOH/DCM to yield 3 (R¹=Boc, R²=allyl) as ayellowish oil (0.25 g). The resulting oil was treated withtrifloroacetic acid in DCM (0.5 M) to yield 33 as yellow film. ¹H NMR(400 MHz, CDCl3) δ 7.93 (s, 1H), 7.84 (s, 1H), 7.33 (s, 1H), 6.71 (d,J=15.6 Hz, 1H), 6.16 (d, J=15.6 Hz, 1H), 5.86 (m, 1H), 5.30 (dd, J=1.3,J=3.8 Hz, 1H), 5.27 (d, J=1.2 Hz, 1H), 5.22 (dd,, J=1.1, 10.2 Hz, 1H),4.38 (s, 1H), 3.91 (m, 2H), 3.69 (m, 2H), 3.58 (m, 2H), 3.30 (d, J=10.5Hz, 1H), 2.98 (d, J=10.5 Hz, 1H), 2.68 (m, 2H), 2.30-2.48 (m, 3H), 1.95(m, 1H), 1.86 (d, J=9.1 Hz, 1H), 1.70-1.81 (m, 6H), 1.49-1.59 (m, 4H),1.45 (d, J=6.6 Hz, 3H). ESI (MH⁺) m/z 566.

Example 31

[0280]

[0281] A sample of compound 33 (0.067 g, 0.12 mmol) was treated withtetrakis(triphenylphosphine)-plladium (0.012 g, 0.01 mmol) and1,3-dimethyl barbituric acid (0.2 g, 1.28 mmol) in DCM at 35° C.overnight, to yield 34 as a solid (0.006 g). ¹H NMR (400 MHz, CDCl3) δ8.04 (s, 1H), 7.92 (s, 1H), 7.33 (s, 2H), 6.77 (d, J=15.6 Hz, 1H), 6.34(d, J=15.6 Hz, 1H), 4.68 (s, 1H), 3.77 (m, 2H), 3.69 (m, 2H), 3.3.29 (d,J=10 Hz, 1H), 2.97 (d, J=10 Hz, 1H), 2.66 (m, 2H), 2.31-2.50 (m, 3H),1.95 (m, 1H), 1.86 (d,, J=9.1 Hz 1H), 1.70-1.81 (m, 7H), 1.49-1.59 (m,4H), 1.45 (d, J=6.6 Hz, 3H). ESI (MH⁺) m/z 526.

Example 32

[0282]

[0283] The mixture of aldehyde compound from Example 28 (0.1 g, 0.22mmol), hydroxylamine hydrochloride (0.024 g, 0.33 mmol) andtriethylamine (0.045 g, 0.45 mmol) in MeOH (0.2 M solution) stirred atroom temperature for 5.5 h. The reaction was treated with saturatedsodium bicarbonate solution and extracted with ethyl acetate, theorganic layer was washed with brine and dried, concentrated, redissolvedin DCM, pale yellow solid came out, rinsed the solid with more DCM,dried to yield 0.04 g 35 as pale yellow solid. ¹H NMR (400 MHz, DMSO) δ11.2 (s, 1H), 10.55 (s, 1H), 7.80 (s, 1H), 7.41 (d, J=9.0 Hz, 1H), 7.26(d, J=9.0 Hz, 1H), 7.23 (s, 1H), 3.64 (br, 2H), 3.43 (m, 2H), 3.30 (s,3H), 3.21 (d, J=10.0 Hz, 1H), 2.88 (d, J=10.0 Hz, 1H), 2.72 (dd, J=4 Hz,J=13 Hz, 1H), 2.58 (m, 1H), 2.49 (t, J=1.75 Hz, 1H), 2.39 (m, 1H), 2.26(m, 1H), 1.86 (m, 1H), 1.72-1.80 (m, 3H), 1.62 (t, J=7.5 Hz, 2H), 1.49(td, J=3.8, Hz J=14 Hz, 2H), 1.38 (m, 1H), 1.35 (d, J=6.6 Hz, 3H), 1.27(m, 1H). ESI (MH⁺) m/z 464.

Example 33

[0284]

[0285] Synthesized in the same way as in Example 29 with the exceptionof replacing hydroxylamine hydrochloride with methoxyaminehydrochloride. ¹H NMR (400 MHz, DMSO) δ 11.12 (s, 1H), 7.77 (s, 1H),7.42 (d, J=8.3 Hz, 1H), 7.35 (s, 1H), 7.27 (d, J=8.3 Hz, 1H), 3.74 (s,2H), 3.65 (d, J=11.4 Hz, 2H), 3.45 (t, J=11.4 Hz, 2H), 3.31 (s, 3H),3.23 (d, J=10.6 Hz, 1H), 2.89 (d, J=8.8 Hz, 1H), 2.72 (dd, J=4 Hz, J=13Hz, 1 H), 2.59 (m, 1H), 2.49 (br, 1H), 2.39 (m, 1H), 2.28 (m, 2H), 1.80(m, 1H), 1.72-1.80 (m, 3H), 1.66 (br, 3H), 1.50 (br, 1H), 1.40 (br, 1H),1.36 (d, J=6.6 Hz, 3H), 1.26 (m, 1H). ESI (MH⁺) m/z 478.

Example 34

[0286]

[0287] The mixture of aldehyde intermediate from Example 28 (0.10 g,0.22 mmol), 2-amino-ethanethiol hydrochloride (0.062 g, 0.54 mmol) andsodium methoxide (0.078 g, 1.4 mmol) in MeOH (0.2 M solution) wasstirred at room temperature overnight. The reaction was treated withsaturated sodium bicarbonate solution and extracted with ethyl acetate,the organic layer was washed with brine, dried, concentrated andpurified by flash chromatography on silica gel eluted with 10:1:0.1DCM-MeOH—NH₄OH to yield 37 as yellow solid (0.06 g). ¹H NMR (400 MHz,CDCl₃) δ 8.09 (s, 1H), 7.83 (s, 1H), 7.33 (s, 2H), 4.65 (d, J=6.7 Hz,1H), 3.79 (m, 2H), 3.70 (dd, J=2.76 Hz, J=9.7 Hz, 1H), 3.53-3.66 (m,4H), 3.48 (s, 3H), 3.29 (d, J=9.5 Hz, 1H), 3.16 (d, J=9.5 Hz, 1H),2.95-2.98 (m, 2H), 2.82 (m, 1H), 2.82 (m, 1H), 2.67 (m, 4H), 2.35-2.6(br, 2H), 2.20 (m, 2H), 2.09 (m, 1H), 1.86 (m, 2H), 1.60-1.80 (m, 2H),1.2-1.55(m, 3H). ESI (MH⁺) m/z 508.

Example 35

[0288]

[0289] Synthesized in two steps: A mixture of amide 8 (0.150 g, 0.32mmol) and (MeO)₂CHN(Me)₂ was heated to 120° C. for 15 min. After cooledto r.t., the whole was directly loaded onto a column eluted with 15-30%MeOH/DCM mixed with 1-10% NH₄OH to yield the corresponding acylamidineas a yellow solid (0.120 g).

[0290] The above acylamnidine was treated with NH₂NH₂.H₂O (0.030 mL, 0.6mmol) in AcOH (1 mL) at 90° C. for 10 min. After cooling to r.t., thewhole was directly loaded onto a column eluted with 10-30% MeOH/DCMmixed with 1-4% NH₄OH to yield triazole 38 as a white solid (0.050 g).¹H NMR δ (DMSO, 500 MHz): 11.18 (s, 1H), 7.77 (s, 1H), 7.43 (d, J=8.5Hz, 1H), 7.28 (d, J=8.5 Hz, 1H), 3.72 (m, 2H), 3.28 (m, 3H), 3.12 (m,1H), 2.79 (m, 1), 2.71 (m, 1H), 2.55 (m, 2H), 2.37 (m, 1H), 2.23 (m,2H), 2.04 (m, 2H), 1.77 (m, 6H), 1.53 (m, 1H), 1.33 (m, 5H), 1.22 (m,1H). MS (ES): 488 [M+H].

Example 36

[0291]

[0292] The mixture of amide 8 (0.695 g, 1.50 mmol) and POCl₃ (0.42 mL,4.5 mmol) in anhydrous pyridine (14 mL) was heated to 120° C. in asealed vessel for 2 h. The mixture was cooled to r.t., poured intosaturated NaHCO₃ solution and extracted with EtOAc. The organic layerwas separated, washed with brine, dried with anhydrous Na₂SO₄,concentrated by rotary evaporation and purified by flash chromatographyon silica gel with a gradient elution of 5-30% MeOH/DCM to yield thecorresponding nitrile as a yellowish solid (0.400 g). ¹H NMR δ (DMSO,500 MHz): 11.19 (s, 1H), 9.79 (s, 1 H), 7.43 (d, J=8.5 Hz, 1H), 7.28 (d,J=8.5 Hz, 1H), 3.87 (m, 2H), 3.50 (m, 2H), 3.31 (m, 2H), 2.96 (m, 1H),2.75 (m, 1H), 2.61 (m, 1H), 2.54 (m, 1H), 2.51 (m, 1H), 1.96 (m, 1H),1.86 (m, 5H), 1.72 (m, 1H), 1.65 (m, 2H), 1.42 (m, 2H), 1.40 (d, J=6.5Hz, 3H), 1.32 (m, 1H). MS (ES): 446 [M+H].

Example 37

[0293]

[0294] Synthesized in two steps: A mixture of amide 8 (0.630 g, 1.36mmol) and LAH (4.76 mL, 1 M in THF, 4.76 mmol) was heated to 70° C. for1 h. After cooling to r.t., the mixture was poured into dilute NH₄OH andextracted with EtOAc. The organic layer was separated, washed withbrine, dried with anhydrous Na₂SO₄, concentrated by rotary evaporationand purified by flash chromatography on silica gel with a gradientelution of 20-50% MeOH/DCM mixed with 0-10% NH₄OH to yield thecorresponding amine as a solid (0.500 g).

[0295] The above amine (0.060 g, 0.133 mmol) was reacted with (CF₃SO₂)₂O(0.067 mL, 0.4 mmol), TEA (0.084 mL, 0.6 mmol) in DCM (2 mL) at 0° C.for 10 min. The mixture was poured into water, basified with saturatedNaHCO₃ and extracted with EtOAc. The organic layer was separated, washedwith brine, dried with anhydrous Na₂SO₄, concentrated by rotaryevaporation and purified by flash chromatography on silica gel with agradient elution with 0-10% MeOH/EtOAc to yield 40 as a yellowish solid(0.045 g). ¹H NMR δ (DMSO, 500 MHz): 11.26 (s, 1H), 7.81 (s, 1H), 7.46(d, J=8.5 Hz, 1H), 7.30 (d, J=8.5 Hz, 1H), 3.67 (m, 1 H), 3.56 (m, 4H),3.35 (m, 1H), 2.97 (m, 4H), 2.82 (m, 2H), 2.66 (m, 2H), 2.31 (m, 2H),1.99 (m, 1H), 1.80 (m, 4H), 1.68 (m, 1H), 1.4-1.5 (m, 4H), 1.39 (d,J=6.5 Hz, 3H). MS (ES): 582 [M+H].

Example 38

[0296]

[0297] A mixture of ester 5 (0.070 g, 0.15 mmol) and MeLi (1.0 mL, 1.6 Min ether, 1.6 mmol) in THF (1.5 mL) was stirred at r.t. for 30 min. Themixture was poured into with saturated NaHCO₃ and extracted with EtOAc.The organic layer was separated, washed with brine, dried with anhydrousNa₂SO₄, concentrated by rotary evaporation and purified by flashchromatography on silica gel eluted with 20% MeOH/DCM to yield 41 as awhite solid (0.012 g). ¹H NMR δ (DMSO, 500 MHz): 11.21 (s, 1H), 7.78 (s,1H), 7.44 (d, J=8.5 Hz, 1H), 7.29 (d, J=8.5 Hz, 1H), 5.95 (s, 1H), 3.69(m, 2H), 3.47 (m, 2H), 3.32 (m, 2H), 2.99 (m, 1H), 2.76 (m, 1H), 2.63(m, 1H), 2.52 (m, 1H), 2.42 (m, 2H), 1.85 (m, 4H), 1.67 (m, 2H),1.2-1.55 (m, 5H), 1.38 (d, J=6.5 Hz, 3H), 1.08 (m, 6H). MS (ES): 479[M+H].

Example 39

[0298]

[0299] Synthesized in three steps: To a solution oftetrahydro-4H-pyran-4-one (5.00 g, 50 mmol) in THF (80 mL) was addedallylmagnesium bromide (60 mL, 1 M/ether, 60 mmol). After stirring atr.t. for 30 min, the reaction was quenched with aqueous NH₄Cl andextracted with ether. The organic layer was separated, washed withbrine, dried with anhydrous Na₂SO₄, concentrated by rotary evaporationand purified by flash chromatography on silica gel with a gradientelution of 50-60% EtOAc/hexanes to yield the alcohol adduct as a clearoil (2.51 g).

[0300] The obtained alkenyl alcohol (1.00 g, 7 mmol) was stirred withNaIO₄ (3.30 g, 15.4 mmol) and OsO₄(40 mg) in MeOH (15 mL) and H₂O (15mL) for 15 min. The entire mixture was directly loaded onto a column.Elution with EtOAc yielded the corresponding aldehyde as a brownish oil(0.70 g).

[0301] The above aldehyde (0.250 g, 1.7 mmol) was stirred with amine 4(0.309 g, 1 mmol) and NaBH(OAc)₃ (0.856 g, 4 mmol) in ClCH₂CH₂Cl (7 mL)for 1.5 h. The mixture was poured into saturated NaHCO₃ and extractedwith EtOAc. The organic layer was separated, washed with brine, driedwith anhydrous Na₂SO₄, concentrated by rotary evaporation and purifiedby flash chromatography on silica gel with a gradient elution of 10-40%MeOH/DCM with 0-10% NH₄OH added to yield 42 as a brownish solid (0.145g). ¹H NMR δ (DMSO, 500 MHz): 11.22 (s, 1H), 7.79 (s, 1H), 7.44 (d,J=8.5 Hz, 1H), 7.29 (d, J=8.5 Hz, 1H), 5.11 (s, 1H), 3.62 (m, 5H), 3.36(m, 1H), 3.03 (m, 1H), 2.76 (m, 1H), 2.62 (m, 1H), 2.51 (m, 1H), 2.42(m, 1H), 1.87 (m, 1H), 1.65 (m, 2H), 1.25-1.6 (m, 9H), 1.39 (d, J=6.5Hz, 3H). MS (ES): 437 [M+H].

Example 40

[0302]

[0303] The mixture of acid 7 (1 g, 2.15 mmol), N-Boc-cystine methylester (1.06 g, 4.52 mmol), bis(2-oxo-3-oxazolidinyl)phosphinic chloride(1.096 g, 4.30 mmol) and triethylamine (0.456 g, 4.52 mmol) in DCM (0.4M solution) was stirred at room temperature overnight. The reaction wastreated with saturated sodium bicarbonate solution and extracted withethyl acetate, the organic layer was washed with brine and dried,concentrated and purified by flash chromatography on silica gel elutedwith 20:1:0.1 DCM:MeOH:NH₄OH to yield a yellow solid (0.6 g).

[0304] The resulting solid was treated with 20% trifluoroacetic acid inDCM. At the completion of the deprotection, excess reagent and solventwere removed by evaporation. The residue was refluxed in benzene (20 mL)and DCE (5 mL, to aid the solubility) overnight. The reaction wastreated with saturated sodium bicarbonate solution and extracted withethyl acetate, the organic layer was washed with brine and dried,concentrated and purified by flash chromatography on silica gel elutedwith 5% MeOH/DCM to yield 43 as a yellow solid (0.16 g). ¹H NMR (400MHz, CDCl₃) δ 8.1 (br, 1H), 7.90 (s, 1H), 7.32 (s, 2H), 5.24 (t, J=8.0Hz, 1H), 3.80 (m, 6H), 3.53-3.66 (m, 5H), 3.48 (s, 3H), 2.66-2.74 (m,2H), 2.52 (br, 2H), 1.80-2.10 (m, 6H), 1.73 (m, 3H), 1.45 (d, J=6.6 Hz,3H), 0.95 (s, 1H. ESI (MH⁺) m/z 564.

Example 41

[0305]

[0306] A sample of compound from Example 40 (0.2 g, 0.35 mmol) wastreated with activated manganese dioxide (0.15 g, 1.75 mmol) in benzenerefluxing overnight, to yield 44 as yellow solid (0.045 g). ¹H NMR (400MHz, CDCl₃) δ 8.17 (s, 1H), 7.91 (s, 1H), 7.75 (s, 1H), 7.32 (s, 2H),5.24 (t, J=8.0 Hz, 1H), 3.93 (s, 3H), 3.84 (m, 2H), 3.58(m, 2H), 3.20(br, 1H), 2.82 (br, 1H), 2.65 (m, 2H), 2.43 (m, 1H), 2.34 (m, 2H),1.90-2.10 (m, 5 H), 1.80 (m, 2H), 1.57 (br, 4H), 1.45 (d, J=6.6 Hz, 3H).ESI (MH⁺) m/z 562.

Example 42

[0307]

[0308] The mixture of amine 4 (0.107 g, 0.35 mmol),[4-(thiazole-2-carbonyl)-tetrahydro-pyran-4-yl]-acetaldehyde (0.069 g,0.29 mmol, prepared similarly as described previously), and sodiumtriacetoxborohydride (0.245 g, 1.15 mmol) in DCE (0.25 M solution)stirred at room temperature for 2 h. The reaction was treated withsaturated sodium bicarbonate solution and extracted with ethyl acetate,the organic layer was washed with brine and dried, concentrated andpurified by flash chromatography on silica gel eluted with 5% MeOH/DCMto yield 45 as yellow solid (0.09 g). ¹H NMR (400 MHz, CDCl₃) δ 7.9 (d,J=2.2 Hz, 2H), 7.80 (s, 1H), 7.46 (d, J=2.2 Hz, 1H), 7.30 (s, 2H), 3.82(m, 2H), 3.61 (t, J=9.8 Hz, 1H), 3.46 (t, J=9.8 Hz, 1H), 3.04 (d, J=9.8Hz, 1H), 2.46-2.68 (m, 6H), 2.18-2.33 (m, 4H), 1.87 (m, 2H), 1.74 (m,1H), 1.45-1.63 (m, 3H), 1.30 (d, J=6.6 Hz, 3H), 1.02 (m, 1H), 0.56 (br,1H). ESI (MH⁺) m/z 532.

Example 43

[0309]

[0310] A sample of ketone from Example 42 (0.039 g, 0.073 mmol) wastreated with sodium borohydride (0.02 g, 0.5 mmol) in THF for 30 min.The reaction was treated with saturated sodium bicarbonate solution andextracted with ethyl acetate, the organic layer was washed with brineand dried, concentrated and purified by flash chromatography on silicagel eluted with 5%-10% MeOH/DCM to yield 46 as yellow solid (0.005 g).¹H NMR (400 MHz, CDCl₃) δ 7.95 (s, 1H), 7.80 (s, 1H), 7.79 (dd,, J=3.3Hz, J=4.6 Hz 1H), 7.38 (dd, J=3.3 Hz J=6.3 Hz, 1H), 7.35 (s, 1H), 4.91(d, J=4.4 Hz, 1H), 3.85 (m, 2H), 3.75 (q, J=1.0 Hz, 2H), 3.41 (m, 1H),3.46 (t, J=9.8 Hz, 1H), 3.19 (dd, J=4.7 Hz, J=7.4 Hz 2H), 3.0 (m, 1H),2.85 (m, 1H), 2.73 (m, 1H), 2.61 (dd, J=9.0 Hz, J=14.9 Hz, 2H),2.05-2.50 (m, 4H), 1.91 (m, 3H), 1.67 (m, 2H), 1.60 (m, 2H), 1.44 (dd,J=6.7 Hz J=9.4 Hz 3H). ESI (MH⁺) m/z 534.

Example 44

[0311]

[0312] Step 1. LAH (0.64 g, 16.7 mL) was added to a dry THF solution (70mL) containing the ester compound from Example 1 (4.0 g, 8.4 mmol) atroom temperature. The solution was heated at reflux for 2 h. Afterheating, water (0.6 mL) was added followed by a 1 N solution of NaOH(0.6 mL), and a final addition of water (1.2 mL). The resulting solidwas filtered washed with copious amount of dichloromethane. The filtratewas concentrated and used in the next step without purification: ESI(MH⁺) m/z 451.

[0313] Step 2. The resulting alcohol compound from above (0.6 g, 1.33mmol) was dissolved in DMSO/Et₃N (2.5:1, 0.2 M) and was treated withSO₃.pyridine complex (0.85 g, 5.33 mmol) and at room temperature. Afterstirring for 2 h, the mixture was poured into water (60 mL) andextracted with dichloromethane (3×100 mL). The organic layers werewashed with brine, dried over Na₂SO₃, and concentrated to give thealdehyde intermediate. ESI (MH⁺) m/z 449.

[0314] Step 3. Sodium hydride (0.96, 40 mmol) was added to a dry DMF(0.2 M) solution containing triethyl phosphonoacetate (4 mL, 20.0 mmol)at room temperature. After stirring for 10 min., the aldehydeintermediate from above (4.51 g, 10.1 mmol) was added and the mixturewas stirred at room temperature overnight. Excess DMF was removed undervacuum and the remaining residue was taken up in a 10% MeOH/DCMsolution, washed with water, dried with Na₂SO₃ and concentrated. Aportion of this material was purified by using preparative HPLC (C18column, 10%-90% acetonitrile/water gradient). ¹H NMR (400 MHz, MeOD) δ7.79 (s, 1H), 7.39 (d, J=8 Hz, 1H), 7.29 (d, J=8 Hz, 1H), 6.87 (d, J=16Hz, 1H), 6.00 (d, J=16 Hz, 1H), 4.22 (q, J=7 Hz, 2H), 3.89 (d, J=10 Hz,1H), 3.77-3.81 (m, 3H), 3.67 (d, J=15 Hz, 1H), 3.57 (t, J=10 Hz, 3H),3.01-3.18 (m, 4H), 2.78-2.91 (m, 4H), 2.52 (m, 1H), 2.20 (d, J=12 Hz,1H), 2.00 (qn, J=6 Hz, 2H), 1.61-1.85 (m, 9H), 1.49 (d, J=6 Hz, 3H),1.31(t, J=7 Hz, 3H). ESI (MH⁺) m/z 519.

Example 45

[0315]

[0316] DIBAL (2.1 eq) was added to a dry THF solution (0.2 M) containingthe ester from Example 44 (1 equiv.) at 0° C. After stirring for 4 h.the reaction was quenched at 0° C. with methanol and warmed to roomtemperature. The solution was concentrated under reduced pressure andthe remaining residue was purified on silica eluting with 0-20%methanol/dichloromethane gradient: ESI (MH⁺) m/z 475.

[0317] Sodium triacetoxyborohydride (3 equiv.) was added to adichloromethane solution (0.2 M) containing aldehyde intermediate fromabove (1 equiv.) and pyrroline (2 equiv.) at room temperature. Afterstirring overnight the solvent was removed using evaporation, and theremaining residue was purified using preparative HPLC (C18 column,10%-90% acetonitrile/water gradient). ¹H NMR (400 MHz, MeOD,) δ 7.78 (s,1H), 7.40 (d, J=8 Hz, 1H), 7.29 (d, J=8 Hz, 1H), 5.93 (d, J=16 Hz, 1H),5.71 (td, J=7, J=16 Hz, 1H), 3.89 (m, 3H), 3.80 (m, 2H), 3.56-3.69 (m,5H), 3.12-3.19 (m, 4 H), 3.01 (t, J=14 Hz, 1H), 2.75-2.92 (m, 3H), 2.52(dd, J=12, J=14 Hz, 1H), 2.10-2.22 (m, 3H), 2.04 (m, 2H), 1.95 (m, 2H),1.64-1.84 (m, 7H), 1.49 (d, J=7 Hz, 3H); ESI (MH⁺) m/z 530.

Example 46

[0318]

[0319] This compound was prepared in the same way as described inExample 45. (TFA salt). ¹H NMR (400 MHz, MeOD,) δ 7.79 (s, 1H), 7.40 (d,J=8 Hz, 1H), 7.29 (d, J=8 Hz, 1H), 5.95 (d, J=16 Hz, 1H), 5.74 (td, J=7,J=16 Hz, 1H), 3.90 (m, 3H), 3.66-3.80 (m, 7H), 3.40-3.60 (m, 6H),3.10-3.26 (m, 2H), 2.99 (t, J=14 Hz, 1 H), 2.75-2.91 (m, 3H), 2.50 (dd,J=12, J=14 1H), 2.17 (d, J=12 Hz, 1H), 1.96 (m, 2H), 1.62-1.82 (m, 7H).1.48 (d, J=7 Hz, 3H). ESI (MH⁺) m/z 594.

Example 47

[0320]

[0321] This compound was prepared in the same way as described inExample 45. (TFA salt). ¹H NMR (400 MHz, MeOD,) δ 7.79 (s, 1H), 7.40 (d,J=8 Hz, 1H), 7.30 (d, J=8 Hz, 1H), 5.87 (d, J=16 Hz, 1H), 5.66 (td, J=7Hz, J=16 Hz, 1H), 3.93 (m, 3H), 3.60-3.80 (m, 7 H), 3.10-3.21 (m, 2H),3.00 (t, J=14 Hz, 1H), 2.75-2.93 (m, 3H), 2.54 (dd, J=12, J=14 Hz, 1H),2.22 (d, J=12 Hz, 1H), 1.80-1.96 (m, 3H), 1.64-1.79 (m, 7H). 1.59 (d,J=7 Hz, 3H), 1.50 (d, J=7 Hz, 3H). ESI (MH⁺) m/z 547.

Example 48

[0322]

[0323] This compound was prepared in the same way as described inExample 45. (TFA salt). ¹H NMR (400 MHz, MeOD,) δ 7.78 (s, 1H), 7.40 (d,J=8 Hz, 1H), 7.29 (d, J=8 Hz, 1H), 5.87 (d, J=16 Hz, 1H), 5.69 (td, J=7,J=16 Hz, 1H), 3.90 (d, J=10 Hz, 1H), 3.59-3.80 (m, 7H), 3.10-3.21 (m,2H), 3.00 (m, 3H), 2.79-2.92 (m, 3H), 2.53 (dd,J=12,J=14 Hz, 1H), 2.19(d, J=12 Hz, 1H), 1.89-1.96 (m, 3H), 1.64-1.79 (m, 6H). 1.49 (d, J=7 Hz,3H), 1.33 (s, 6H). ESI (MH⁺) m/z 548.

Example 49

[0324]

[0325] This compound was prepared in the same way as described inExample 45. (TFA salt). ¹H NMR (400 MHz, MeOD,) δ 7.78 (s, 1H), 7.40 (d,J=8 Hz, 1H), 7.29 (d, J=8 Hz, 1H), 5.91 (d, J=16 Hz, 1H), 5.69 (td, J=7Hz, J=16 Hz, 1H), 4.03 (q, J=9 Hz, 2H), 3.84-3.91 (m, 3H), 3.78 (m, 2H),3.58-3.70 (m, 3H), 3.10-3.21 (m, 2H), 2.99 (t, J=14 Hz, 1H), 2.77-2.91(m, 3H), 2.53 (dd, J=12, J=14 Hz, 1 H), 2.19 (d, J=12 Hz, 1H), 1.78-1.97(m, 3H), 1.57-1.78 (m, 6H). 1.48 (d, J=7 Hz, 3H). ESI (MH⁺) m/z 558.

Example 50

[0326]

[0327] A sample of the aldehyde intermediate obtained in Step 1 inExample 44 was treated methyl Grinard reagent (2.5 equiv.) in dry THF(0.2 M) at room temperature. After stirring overnight the solvent wasremoved using evaporation, and the remaining residue was purified usingpreparative HPLC (C18 column, 10%-90% acetonitrile/water gradient). (TFAsalt). ¹H NMR (400 MHz, MeOD,) δ 7.79 (s, 1H), 7.40 (d, J=8 Hz, 1H),7.29 (d, J=8 Hz, 1H), 5.62 (ddd, J=2, J=6, J=16 Hz, 1H), 5.51 (d, J=16Hz, 1H), 4.33 (qn, J=6 Hz, 1H), 3.89 (d, J=12 Hz, 1H) 3.77 (m, 2H),3.59-3.69 (m, 3H), 3.10-3.21 (m, 2H), 3.00 (t, J=14 Hz, 1H), 2.77-2.92(m, 3H), 2.52 (dd, J=12, J=14 Hz, 1H), 2.20 (d, J=12 Hz, 1H), 1.83-1.90(m, 3 H), 1.59-1.74 (m, 6H). 1.50 (d, J=7 Hz, 3H), 1.30 (d, J=7 Hz, 3H).ESI (MH⁺) m/z 491.

Example 51

[0328]

[0329] obtained from treatment of aldehyde intermediate from Step 1 inExample 44 with an isopropyl Grinard. (TFA salt). ¹H NMR (400 MHz,MeOD,) δ 7.79 (s, 1H), 7.40 (d, J=8 Hz, 1H), 7.29 (d, J=8 Hz, 1H),5.43-5.60 (m, 2H), 3.88 (m, 2H), 3.77 (m, 2H), 3.59-3.69 (m, 3H),3.10-3.21 (m, 2H), 3.01 (t, J=14 Hz, 1H). 2.80-2.92 (m, 3H), 2.52 (dd,J=12, J=14 Hz, 1H), 2.20 (d, J=12 Hz, 1H), 1.63-1.93 (m, 10H), 1.49 (d,J=7 Hz, 3H), 0.97 (d, J=6 Hz, 3H), 0.94 (d, J=6 Hz, 3H). ESI (MH⁺) m/z519.

Example 52

[0330]

[0331] Obtained from treatment of aldehyde intermediate from Step 1 inExample 44 with a t-butyl Grinard. (TFA salt). ¹H NMR (400 MHz, MeOD,) δ7.79 (s, 1H), 7.40 (d, J=8 Hz, 1H), 7.29 (d, J=8 Hz, 1H), 5.65 (dd, J=7,J=16 Hz, 1H), 5.51 (d, J=16 Hz, 1H), 3.85 (m, 1H), 3.78 (m, 3H),3.59-3.69 (m, 3H), 3.10-3.21 (m, 2H), 3.02 (t, J=14 Hz, 1H), 2.80-2.93(m, 3H), 2.53 (dd, J=12, J=14 Hz, 1H), 2.19 (d, J=12 Hz, 1H), 1.80-1.90(m, 3H), 1.62-1.77 (m. 6H), 1.50 (d, J=7 Hz, 3H), 0.96 (s, 9 H). ESI(MH⁺) m/z 533.

Example 53

[0332]

[0333] Lithium hydroxide (30 mg, 1.3 mmol) was added to a THF/watersolution (1:1, 0.2 M) containing ester intermediate from Example 44(0.54 g, 1.0 mmol) and heated at reflux for 3 h. After cooling to roomtemperature, the solution was titrated with a 3 N HCl solution toneutral pH and concentrated to dryness using reduced pressure. Thismaterial was used in the next step without purification. ESI (MH⁺) m/z491.

[0334] General synthesis for analogs amide formation. HBTU (3 equiv.)was added to a dichloromethane solution (0.2 M) containing,triethylamine (3 equiv.), acid intermediate from above(1 equiv.), andthe respective amine (2 equiv.) at room temperature. After stirringovernight the solvent was removed using evaporation, and the remainingresidue was purified using preparative HPLC (C18 column, 10%-90%acetonitrile/water gradient).

[0335] R=Et, (TFA Salt). ¹H NMR (400 MHz, MeOD,) δ 7.79 (s, 1H), 7.40(d, J=8 Hz, 1H), 7.29 (d, J=8 Hz, 1H), 6.62 (d, J=16 Hz, 1H), 6.03 (d,J=16 Hz, 1H), 3.89 (d, J=10 Hz, 1H), 3.79(m, 2H), 3.50-3.73 (m, 4H),3.13-3.21 (m, 2H), 3.02 (m, 1H), 2.80-3.0 (m, 3H), 2.53 (dd, J=12, J=14Hz, 1H), 2.20 (d, J=12 Hz, 1H), 1.90-2.0 (m, 3H), 1.60-1.87 (m. 7H),1.50 (m, 4H), 1.57 (t, J=7 Hz, 3H). ESI (MH⁺) m/z 518.5.

Example 54

[0336]

[0337] (TFA salt). ¹H NMR (400 MHz, MeOD,) δ 7.79 (s, 1 H), 7.40 (d, J=8Hz, 1H), 7.29 (d, J=8 Hz, 1H), 6.63 (d, J=16 Hz, 1H), 6.02 (d, J=16 Hz,1H), 4.06 (qn, J=7 Hz, 1H), 3.84 (d, J=10 Hz, 1H), 3.70 (m, 2H),3.56-3.69 (m, 3H), 3.10-3.19 (m, 2H), 3.02 (t, J=14 Hz, 1H), 2.80-2.93(m, 3H), 2.52 (dd, J=12, J=14 Hz, 1H), 2.19 (d, J=12 Hz, 1H), 1.62-1.99(m. 9H), 1.50 (d, J=7 Hz, 3H), 1.19 (d, J=7 Hz, 6H). ESI (MH⁺) m/z532.5.

Example 55

[0338]

[0339] (TFA salt) ¹H NMR (400 MHz, MeOD,) δ 7.76 (s, 1H), 7.35 (d, J=8Hz, 1H), 7.24 (d, J=8 Hz, 1H), 6.70 (d, J=16 Hz, 1H), 6.07 (d, J=16 Hz,1H), 3.76 (m, 2H), 3.58 (t, J=9 Hz, 2H), 3.40 (s, 2H), 3.31 (m, 1 H),3.02 (d, J=10 Hz, 1H), 2.75 (dd, J=3 Hz, J=16 Hz, 1H), 2.64 (qn, J=7 Hz,1H), 2.40 (m, 3H), 2.04 (t, J=10 Hz, 1H), 1.69-1.93 (m, 9H), 1.63 (m,6H), 1.49 (m, 2H), 1.43 (d, J=7 Hz, 3H), 1.34 (m, 1H). ESI (MH⁺)m/z 588.

Example 56

[0340]

[0341] Tosylmethyl isocyanide (22 mg, 0.1 mmol) was added to a methanolsolution (5 mL) containing potassium carbonate (19 mg, 1.2 mmol) andaldehyde intermediate from Step 2 of Example 44 (50 mg, 0.1 mmol). Thesolution was heated at reflux for 24 h. After cooling, the product waspurified using preparative HPLC (C18 column, 10%-90% acetonitrile/watergradient): ¹H NMR (400 MHz, MeOD,) δ 8.23 (s, 1H), 7.75 (s, 1H), 7.39(d, J=8 Hz, 1H), 7.25 (d, J=8 Hz, 1H), 7.11 (s, 1H), 7.11 (s, 1H), 3.81(d, J=12 Hz, 3H), 3.59 (d, J=12 Hz, 1H), 3.45 (t, J=11 Hz, 2H),2.72-3.06 (m, 6H), 2.47(dd, J=12, J=14 Hz, 1H), 2.13 (m, 4H), 1.77-1.88(m, 3H), 1.55-1.67 (m, 2H), 1.44 (d, J=7 Hz, 3H). ESI (MH⁺) m/z 488.

Example 57

[0342]

[0343] To a mixture of acid 12 (0.150 g, 0.32 mmol), DMF (2 drops) andDCM (4 mL) was added (COCl)₂ (1.0 mL, 2 M in DCM, 7.2 mmol). When thegas release ceased, the mixture was placed under high vacuum to obtain asolid. To this solid was added DCM (4 mL) and MeNH₂ (8 mL, 2 M/THF, 16mmol). The mixture was stirred for 1 h at r.t., poured into brine andextracted with EtOAc. The organic layer was separated, washed withbrine, dried with anhydrous Na₂SO₄, concentrated by rotary evaporationand purified by flash chromatography on silica gel eluted with agradient elution of 20-40% MeOH/DCM mixed with 0-7% NH₄OH to yield 60 asa yellowish solid (0.095 g). MS (ES): 476 [M+H].

Example 58

[0344]

[0345] A mixture of acid 12 (0.926 g, 2 mmol), glycinamide HCl (0.442 g,4 mmol), EDC.HCl (0.960 g, 5 mmol), HOBt (0.676 g, 5 mmol), NMP (2.0 mL,18 mmol), DCM (10 mL) and DMF (10 mL) was stirred at r.t. for 3 h. Themixture was poured into saturated NaHCO₃ and extracted with EtOAc. Theorganic layer was separated, washed with brine, dried with anhydrousNa₂SO₄, concentrated by rotary evaporation and purified by flashchromatography on silica gel with a gradient elution of 20-40% MeOH/DCMmixed with 1-8% NH₄OH to yield 61 as a brownish solid (0.650 g). ¹H NMRδ (DMSO, 500 MHz): 11.19 (s, 1H), 7.89 (t, J=7.5 Hz, 1H), 7.78 (s, 1H),7.43 (d, J=8.5 Hz, 1H), 7.28 (d, J=8.5 Hz, 1H), 7.20 (s, 1H), 6.97 (s,1H), 3.66 (m, 4H), 3.39 (m, 2H), 3.24 (m, 1H), 3.29 (m, 1H), 2.74 (m,1H), 2.60 (m, 1H), 2.40 (m, 1H), 2.28 (m, 2H), 2.12 (m, 1H), 2.03 (m,2H), 1.89 (m, 1H), 1.80 (m, 1H), 1.71 (m, 2H), 1.63 (m, 1H), 1.42 (m,5H), 1.38 (d, J=6.5 Hz, 3H), 1.27 (m, 1). MS (ES): 519 [M+H].

Example 59

[0346]

[0347] The corresponding amine compound was prepared following the sameprocedures as depicted for Example 15, with the exception ofsubstituting 4-allyl-tetrahydropyran carboxylic acid methyl ester with1-allyl-1-cyclohexyl carboxylic acid methyl ester.

[0348] A mixture of this amine (0.235 g, 0.54 mmol), EtSO₂Cl (0.139 g,1.08 mmol), TEA (0.109 g, 1.08 mmol) in DCM (5 mL) was stirred at r.t.for 45 min. The mixture was poured into saturated NaHCO₃ and extractedwith EtOAc. The organic layer was separated, washed with brine, driedwith anhydrous Na₂SO₄, concentrated by rotary evaporation and purifiedby flash chromatography on silica gel with a gradient elution of 20 %MeOH/DCM mixed with 0-1% NH₄OH to yield 62 as a solid (0.037 g). ¹H NMRδ (DMSO, 500 MHz): 11.20 (s, 1H), 7.79 (s, 1H), 7.44 (d, J=8.5 Hz, 1H),7.29 (d, J=8.5 Hz, 1H), 7.16 (s, 1H), 3.33 (m, 2H), 3.00 (m, 3H), 2.74(m, 1H), 2.63 (m, 1H), 2.37-2.55 (m, 4H), 1.95 (m, 1H), 1.84 (m, 6H),1.72 (m, 1H), 1.55 (m, 2H), 1.40 (m, 6H), 1.39 (d, J=6.5 Hz, 3H), 1.25(t, J=7.2 Hz, 3 H). MS (ES): 526 [M+H].

Example 60

[0349]

[0350] A mixture of amine intermediate from Example 59 (0.056 g, 0.13mmol), CF₃CH₂SO₂Cl (0.036 g, 0.15 mmol), TEA (0.042 mL, 0.3 mmol) in DCM(1 mL) was stirred at r.t. for 15 min. The mixture was poured intosaturated NaHCO₃ and extracted with EtOAc. The organic layer wasseparated, washed with brine, dried with anhydrous Na₂SO₄, concentratedby rotary evaporation and purified by flash chromatography on silica gelwith a gradient elution of 20% MeOH/DCM mixed with 0-1% NH₄OH to yield63 as a solid (0.030 g). ¹H NMR δ (DMSO, 500 MHz): 11.20 (s, 1 H), 8.19(s, 1H), 7.79 (s, 1H), 7.44 (d, J=8.5 Hz, 1H), 7.28 (d, J=8.5 Hz, 1H),4.31 (m, 2H), 3.28 (m, 2H), 2.97 (m, 1H), 2.75 (m, 1H), 2.63 (m, 1H),2.48 (m, 2H), 2.37 (m, 1H), 1.95 (m, 1H), 1.84 (m, 5H), 1.74 (m, 1H),1.5-1.68 (m, 5H), 1.2-1.5 (m, 6H), 1.38 (d, J=6.5 Hz, 3H). MS (ES): 580[M+H].

Example 61

[0351]

[0352] A mixture of amine intermediate from Example 59 (0.097 g, 0.224mmol), 4,4-dioxo-tetrahydrothiopyranyl carboxylic acid (0.040 g, 0.224mmol), EDC.HCl (0.107 g, 0.56 mmol), HOBt (0.076 g, 0.56 mmol), NMP(0.275 mL, 2.5 mmol), DCM (1.5 mL) and DMF (1.5 mL) was stirred at r.t.overnight. The mixture was poured into saturated NaHCO₃ and extractedwith EtOAc. The organic layer was separated, washed with brine, driedwith anhydrous Na₂SO₄, concentrated by rotary evaporation and purifiedby flash chromatography on silica gel with a gradient elution of 10-20%MeOH/DCM mixed with 0-3% NH₄OH to yield 64 as a white solid (0.023 g).¹H NMR δ (DMSO, 500 MHz): 11.25 (s, 1H), 7.79 (s, 1H), 7.45 (d, J=8.5Hz, 1H), 7.29 (d, J=8.5 Hz, 1H), 3.13 (m, 5H), 2.78 (m, 2H), 2.64 (m,1H), 2.55 (m, 1H), 2.43 (m, 2H), 2.08 (m, 1OH), 1.2-1.55 (m, 16H). MS(ES): 594 [M+H].

Example 62

[0353]

[0354] Synthesized according to the same sequence as used for thesynthesis of compound in Example 4. ¹H NMR δ (DMSO, 500 MHz): 11.20 (s,1H), 7.78 (s, 1H), 7.44 (d, J=8.5 Hz, 1H), 7.29 (d, J=8.5 Hz, 1H), 3.58(s, 1H), 3.30 (m, 2H), 2.97 (m, 1H), 2.75 (m, 1H), 2.61 (m, 1H), 2.39(m, 3H), 2.02 (m, 3H), 1.80 (m, 4H), 1.59 (m, 4H), 1.45 (m, 3H), 1.38(d, J=6.5 Hz, 3H), 1.32 (m, 1H). MS (ES): 449 [M+H].

Example 63

[0355]

[0356] To a mixture of acid 65 (0.070 g, 0.15 mmol), DMF (1 drop) andDCM (2 mL) was added (COCl)₂ (0.5 mL, 2 M in DCM, 1 mmol). When the gasrelease ceased, the mixture was placed under high vacuum to obtain asolid. To this solid was added DCM (2 mL), MeNH₂(2 mL, 2 M in THF, 2mmol). The mixture was stirred at r.t. for 1 h, poured into saturatedNaHCO₃ and extracted with EtOAc. The organic layer was separated, washedwith brine, dried with anhydrous Na₂SO₄, concentrated by rotaryevaporation and purified by flash chromatography on silica gel elutedwith a gradient elution of 5-30% MeOH/DCM mixed with 0-5% NH₄OH to yield66 as a white solid (0.060 g). ¹H NMR δ (DMSO, 500 MHz): 11.21 (s, 1H),7.79 (s, 1H), 7.59 (s, 1H), 7.44 (d, J=8.5 Hz, 1H), 7.29 (d, J=8.5 Hz,1H), 2.76 (m, 1H), 2.55 (m, 1H), 2.60 (d, J=4.0 Hz, 3H), 2.41 (m, 2H),2.19 (m, 1H), 2.01 (m, 2H), 1.81 (m, 4H), 1.33-1.58 (m, 1OH), 1.39 (d,J=6.5 Hz, 3H). MS (ES): 462 [M+H].

Example 64

[0357]

[0358] A mixture of acid 65 (0.100 g, 0.22 mmol),2-aminomethyl-2-propanol (0.120 g, 0.67 mmol), EDC.HCl (0.127 g, 0.66mmol), HOBt (0.089 g, 0.66 mmol), NMP (0.25 mL, 2.3 mmol), DCM (2 mL)and DMF (2 mL) was stirred at r.t. overnight. The mixture was pouredinto saturated NaHCO₃ and extracted with EtOAc. The organic layer wasseparated, washed with brine, dried with anhydrous Na₂SO₄, concentratedby rotary evaporation and purified by flash chromatography on silica gelwith a gradient elution of 10-20% MeOH/DCM mixed with 1-2% NH₄OH toyield 67 as a white solid (0.100 g). MS (ES): 520 [M+H].

Example 65

[0359]

[0360] Synthesized according to the same sequence as was used for thesynthesis of compound 12 (Example 9). ¹H NMR δ (DMSO, 500 MHz): 11.21(s, 1H), 7.78 (s, 1H), 7.44 (d, J=8.5 Hz, 1H), 7.29 (d, J=8.5 Hz, 1H),3.31 (m, 2H), 2.99 (m, 1H), 2.75 (m, 1H), 2.61 (m, 1H), 2.37 (m, 6H),1.8-2.05 (m, 8H), 1.25-1.50 (m, 6H). MS (ES): 435 [M+H].

Example 66

[0361]

[0362] Synthesized according to the same sequence as was used for thesynthesis of example 2 ¹H NMR (400 MHz, CD₃OD) δ 7.78 (s, 1 H), 7.39 (d,J=8.0 Hz, 1H), 7.27 (d, J=8.0 Hz, 1H), 3.75 (m, 1H), 3.40 (m, 1H), 3.14(m, 2H), 3.00 (m, 2H), 2.60(m, 1H), 2.40-2.50 (m, 4H), 1.50-2.00 (m, 4H)1.44 (d, J=6.4 Hz, 3H), 1.22 (s, 6H). ESI (MH⁺) m/z 423.

Example 67

[0363]

[0364] Compound 69 from Example 66 (1.03 g, 2.43 mmol) in 5 mL DCM wastreated with oxalyl chloride (2.1 mL, 24.3 mmol) and two drops of dryDMF. After 30 min., the reaction was brought to dryness under lowpressure. DCM (15 mL) was added and the flask was put into an ice bath.Much excess ammonia in dry DCM was added slowly to react with thepreviously formed carbonyl chloride. In around 20 min., the reactionshowed completion by LC-MS. Water (30 mL) was added and the aqueouslayer was extracted with DCM (3×20 mL). The organic layers werecombined, dried and concentrated. Purification by flash chromatographyon silica gel with 5-10% MeOH/DCM afforded 70 (800 mg, 1.90 mmol) aslight brown film. ¹H NMR (400 MHz, CD₃OD) δ 7.78 (s, 1H), 7.39 (d, J=8.0Hz, 1H), 7.26 (d, J=8.0 Hz, 1H), 3.77 (m, 1H), 3.41 (m, 1H), 2.98 (m,2H), 2.55(m, 1H), 2.10-2.50 (m, 4H), 1.45-2.00 (m, 4H) 1.41 (d, J=6.4Hz, 3H), 1.21 (s, 6H). ESI (MH⁺) m/z 422.

Example 68

[0365]

[0366] The mixture of the acid 69 (Example 66) (50 mg, 0.118 mmol), EDC(68 mg, 0.354 mmol), HOBT (16 mg, 0.118 mmol), NMM (0.039 mL, 0.354mmol) and excess ethylamine (˜10 equiv.) in 2 mL DCM was stirred at roomtemperature overnight. The solvent was removed under vacuum and 1 mL DMFand 0.2 mL water were added. The solution was injected directly to HPLC(reverse phase) to render 30 mg (0.066 mmol) of 71 as yellow film. ¹HNMR (400 MHz, CD₃OD) δ 7.77 (s, 1H), 7.37 (d, J=8.4 Hz, 1H), 7.26 (d,J=8.4 Hz, 1H), 3.30 (m, 2H), 3.21 (q, J=7.2 Hz, 1H), 3.10-3.20 (m, 4H),2.55(m, 1H), 2.15-2.50 (m, 4H), 1.50-2.00 (m, 4.44 (d, J=6.4 Hz, 3H),1.22 (s, 6H), 1.13 (t, J=7.2 Hz, 3H). ESI (MH⁺) m/z 450.

Example 69

[0367]

[0368] Compound 72 was prepared following same procedures as describedfor Example 67. ¹H NMR (400 MHz, CD₃OD) 6 7.79 (s, 1H), 7.39 (d, J=8.4Hz, 1H), 7.28 (d, J=8.4 Hz, 1H), 7.07 (d, J=8.4 Hz, 2H), 6.75 (d, J=8.4Hz, 2H), 3.82 (m, 1H), 3.61 (m, 1H), 3.20 (m, 2H), 2.65-3.00 (m, 6H),2.50 (m, 1H), 2.15 (m, 3H),1.60-1.90 (m, 3H) 1.45 (d, J=6.4 Hz, 3H). ESI(MH⁺) m/z 443.

Example 70

[0369]

[0370] The same procedure was followed as for compound 71 (Example 68).¹H NMR (400 MHz, CD₃OD) δ 7.78 (s, 1 H), 7.39 (d, J=8.4 Hz, 1H), 7.29(d, J=8.4 Hz, 1H), 3.90 (m, 1H), 3.70 (m, 1H), 3.5 (m, 2H), 3.30 (m,3H), 2.95 (s, 6H), 2.50-2.90 (m, 5H), 2.50 (m, 1H), 2.00-2.20 (m, 4H),1.60-1.90 (m, 2H) 1.49 (d, J=6.8 Hz, 3H), 1.27 (s, 6H). ESI (MH⁺) m/z493.

Example 71

[0371]

[0372] The same procedure was followed as for compound 71 (Example 68).¹H NMR (400 MHz, CD₃OD) 6 7.77 (s, 1 H), 7.38 (d, J=8.4 Hz, 1H), 7.27(d, J=8.4 Hz, 1H), 7.20-7.40 (m, 5H), 3.90 (m, 1H), 3.70 (br, 2H),3.30-3.65 (m, 8H), 3.20 (m, 2H), 2.50-2.90 (m, 5H), 2.50 (m, 1H),2.00-2.20 (m, 3H), 1.60-1.90 (m, 3 H) 1.49 (d, J=6.8 Hz, 3H), 1.27 (s,6H). ESI (MH⁺) m/z 582.

Example 72

[0373]

[0374] The same procedure was followed as for compound 71 (Example 68).¹H NMR (400 MHz, CD₃OD) δ 7.77 (s, 1 H), 7.38 (d, J=8.4 Hz, 1H), 7.27(d, J=8.4 Hz, 1H), 7.20-7.40 (m, 5H), 3.88 (m, 1 H), 3.69 (br, 2H),3.30-3.65 (m, 5 H), 3.21 (m, 2H), 2.50-2.90 (m, 5H), 2.50 (m, 1H),2.00-2.20 (m, 4H), 1.60-1.90 (m, 6H) 1.48 (d, J=6.8 Hz, 3H), 1.26 (s,6H). ESI (MH⁺) m/z 596.

Example 73

[0375]

[0376] Compound 76 was prepared by reducing compound 69 and oxidizingthe corresponding alcohol with SO₃.pyridine in DMSO/TEA as described inExample 44. To the solution of compound 76 (25 mg, 0.061 mmol) in 2 mLDCM was added NaBH(OAc)₃ (26.0 mg, 0.122 mmol) followed by the additionof amine (11 mg, 0.122 mmol). The reaction mixture was kept stirring forovernight. The solvent was removed and 1 mL DMF was added to dissolvethe mixture. After filtration, the solution was injected directly toreverse HPLC to render pure yellow film 77 (18 mg, 0.038 mmol) as theproduct. ¹H NMR (400 MHz, CD₃OD) δ 7.78 (s, 1 H), 7.40 (d, J=8.4 Hz,1H), 7.30 (d, J=8.4 Hz, 1 H), 3.93 (m, 1 H), 3.68 (m, 1 H), 3.60 (s, 2H), 3.30-3.40 (m, 4 H),2.75-3.10(m,4H), 2.50(m, 1 H),2.00-2.20(m,3H),1.60-1.90(m,3H) 1.49 (d, J=6.4 Hz, 3 H), 1.38 (s, 6 H), 1.14 (s, 6 H).ESI (MH⁺) m/z 480.

Example 74

[0377]

[0378] Compound 78 was obtained in several steps following a sequencesimilar to that of Example 10. At 0° C., to the solution of compound 78(6 mg, 0.015 mmol) and triethylamine (6 mg, 0.06 mmol), was addedmethansulfonic anhydride (7.8 mg, 0.045 mmol). The reaction was keptstirring for overnight. The solvent was removed under vacuum and theresulted residue was subjected to reverse HPLC to render pure yellowsolid 79 (2.0 mg, 0.004 mmol). ¹H NMR (400 MHz, CD3OD) δ 7.78 (s, 1 H),7.40 (d, J=8.4 Hz, 1 H), 7.30 (d, J=8.4 Hz, 1 H), 3.50-3.70 (m, 2 H),3.00-3.30 (m, 3 H), 2.70 (s, 3 H), 2.60-3.00 (m, 4 H), 1.60-2.20 (m, 6H), 1.47 (d, J=6.4 Hz, 3 H), 1.20 (s, 6 H). ESI (MH⁺) m/z 472.

Example 75

[0379]

[0380] A mixture of amine 4 (0.9 g, 2.92 mmol), sodiumtriacetoxborohydride (2.5 g, 11.7 mmol) and aldehyde 80 (1.0 g, 3.5mmol) in DCE (0.25 M solution) was stirred at room temperature for 3 h.The reaction mixture was treated with saturated sodium bicarbonatesolution and extracted with ethyl acetate, the organic layer was washedwith brine and dried, concentrated and purified by flash chromatographyon silica gel eluted with 3% MeOH/DCM to yield 81 as yellow solid (0.73g). Compound 81 was treated with trifloroacetic acid in DCM for 0.5 h,concentrated and purified by flash chromatography on silica gel elutedwith 20:1:0.1 DCM:MeOH:NH₄OH to yield 82 as pale yellow solid (0.49 g).¹H NMR (400 MHz, DMSO) δ 11.2 (s, 1 H), 8.03 (s, 1 H), 7.41 (d, J=8.4Hz, 1 H), 7.25 (d, J=8.4 Hz, 1 H), 3.64 (s, 3 H), 3.2 (dd, J=2.4 Hz,J=9.2 Hz, 1 H), 2.86 (m, 2 H), 2.84 (dd, J=2.4 Hz J=9.2 Hz, 1 H),2.50-2.75 (m, 4 H), 2.10-2.45 (m, 4 H), 2.0 (d, J=12 Hz ,2 H), 1.55-1.90(m, 5 H), 1.47 (m, 2 H), 1.35-1.4 (m, 2H), 1.35 (d, J=6.6 Hz, 3 H), 1.25(m, 1 H). ESI (MH⁺) m/z 478.

[0381] Compound 82 (0.1 g, 0.21 mmol) was treated with formaldehyde(0.01 g, 0.3 mmol) and sodium triacetoxborohydride (0.178 g, 0.84 mmol)in DCE for 3 h. at room temperature. The reaction was treated withsaturated sodium bicarbonate solution and extracted with ethyl acetate,the organic layer was washed with brine and dried, concentrated andpurified by flash chromatography on silica gel eluted with 20:1:0.1DCM:MeOH:NH₄OH to yield 83 as yellow solid (0.01 g). ¹H NMR (400 MHz,CDCl₃) δ 7.93 (s, 1 H), 7.84 (s, 1 H), 7.33 (s, 2 H), 3.71 (s, 3 H), 3.4(d, J=6.4 Hz, 1 H), 2.96 (d, J=6.4 Hz, 1 H), 2.67 (m, 4 H), 2.47 (dd,J=8.0 Hz J=12 Hz, 1 H), 2.10-2.40 (m, 7 H), 1.45-1.95 (m, 11 H), 1.35(d, J=6.6 Hz, 3 H). ESI (MH+) m/z 492.

[0382] Compound 82 (0.05 g, 0.105 mmol) was treated with mesyl chloride(0.012 g, 0.105 mmol) and triethylamine (0.013 g, 0.013 mmol) in DCM for2 h. at 0° C. The reaction was treated with saturated sodium bicarbonatesolution and extracted with ethyl acetate, the organic layer was washedwith brine and dried, concentrated and purified by flash chromatographyon silica gel eluted with 20:1:0.1 DCM:MeOH:NH4OH to yield 84 as yellowsolid (0.055 g). ¹H NMR (400 MHz, CDCl3) δ 8.10 (s, 1 H), 7.86 (s, 1 H),7.32 (s, 2 H), 3.74 (s, 3 H), 3.63 (d, J=9.5 Hz, 1 H), 2.93 (d, J=9.5Hz, 1 H), 2.60-2.80 (m, 7 H), 2.20-2.50 (m, 5 H), 1.96 (t, J=10.8 Hz, 1H), 1.8 (m, 4 H), 1.73 (t, J=7.8, 1 H), 1.60 (t, J=12 Hz, 2 H),1.30-1.50 (m, 3H), 1.35 (d, J=6.6 Hz, 3 H). ESI (MH⁺) m/z 556.

Example 76

[0383]

[0384] Comound 83 (0.02 g, 0.04 mmol) was treated with lithiumaluminumhydride (0.02 g, 0.042 mmol) in THF for 2 h. at roomtemperature. The reaction was treated with saturated sodium bicarbonatesolution and extracted with ethyl acetate, the organic layer was washedwith brine and dried, concentrated and purified by flash chromatographyon silica gel eluted with 20:1:0.1 DCM:MeOH:NH₄OH to yield 85 as yellowsolid (0.055 g). ¹H NMR (400 MHz, CDCl₃) δ 8.1 (s, 1 H), 7.80 (s,l H),7.33 (s, 2 H), 3.41 (s, 3 H), 3.15 (d, J=10.5 Hz, 1 H), 2.35-2.65 (m, 7H), 2.67 (m, 4 H), 2.05 (m, 2 H), 1.5-1.9 (m, 12 H), 1.40 (d, J=6.6 Hz,3 H). ESI (MH⁺) m/z 464.

Example 77

[0385]

[0386] Compound 83 (4 g, 8.15 mmol) was treated with lithium hydroxide(0.391 g, 16.3 mmol) in THF, MeOH and water reflux overnight. Thereaction was treated with 1 N HCl solution and extracted with isopropylalcohol and chloroform, the organic layer was washed with brine anddried, concentrated and purified by flash chromatography on silica geleluted with 10:1:0.1 DCM:MeOH:HOAc to yield 2 g pink powder (acid). Theacid (0.052 g, 0.11 mmol) was reacted with 2-methoxyethylamine (0.0098g, 0.13 mmol), under the condition of EDC (0.063 g, 0.33 mmol), HOBt(0.0147 g, 0.11 mmol), and NMM (0.033 g, 0.33 mmol) in DMF (1 mL) atroom temperature overnight. The reaction was treated with saturatedsodium bicarbonate solution and extracted with ethyl acetate, theorganic layer was washed with brine and dried, concentrated and purifiedby flash chromatography on silica gel eluted with 20:1:0.1DCM:MeOH:NH₄OH to yield 86 as brown oil (0.003 g). ¹H NMR (400 MHz,CDCl₃) δ 8.05 (s, 1H), 7.83 (s, 1H), 7.34 (s, 2H), 3.48 (s, 3H), 3.35(s, 3H), 3.05 (d, J=11.2 Hz, 1H), 2.88 (m, 2H), 2.60-2.75 (m, 3H),2.35-2.55 (m, 9H), 2.05 (m, 2H), 1.48-1.60 (m, 5H), 1.22 (s, 6H), 0.85(m, 2H). ESI (MH⁺) m/z 464.

Example 78

[0387]

[0388] Intermediate 88. EDC (1.12 g, 5.9 mmol) was added to adichloromethane solution (0.2 M) containing, triethylamine (1 mL.),amine 4 (1.2 g, 3.9 mmol), and the carboxylic acid 87 (0.97 g, 3.9 mmol)at room temperature. After stirring overnight the mixture was washedwith water, dried over Na₂SO₄, and concentrated to give intermediate 88.This material was used in the next step without purification. ESI (MH⁺)m/z 539.

[0389] Analog 89 (TFA Salt). LAH (200 mg) was added to a dry THFsolution (10 mL) containing 88 (2.0 g, 3.9 mmol) at room temperature.The solution was heated at reflux for 2 h. After heating, water (0.2 mL)was added followed by a 1 N solution of NaOH (0.2 mL), and a finaladdition of water (0.4 mL). The resulting solid was filtered washed withcopious amounts of dichloromethane. A portion of this material waspurified by using preparative HPLC (C18 column, 10%-90%acetonitrile/water 15 gradient). ¹H NMR (400 MHz, MeOD) 6 7.79 (s, 1H),7.36-7.42 (m, 5H), 7.25-7.32 (m, 2H), 4.14 (d, J=13 Hz, 1H), 3.90 (t,J=11 Hz, 1H), 3.67 (d, J=11 Hz, 1H) 3.32-3.52 (m, 3H), 2.81-3.14 (m,8H), 2.53 (m, 2H), 2.19-2.31 (m, 3H), 1.66-1.89 (m, 3H), 1.50 (m, 3H).ESI (MH⁺) m/z 511.

[0390] General synthesis for analogs 90 and 91. Sodiumtriacetoxyborohydride (3 equiv.) was added to a dichloromethane solution(0.2 M) containing amine 89 (1 equiv.) and the respective ketone oraldehyde (3 equiv.) at room temperature. After stirring overnight thesolvent was removed using evaporation, and the remaining residue waspurified using preparative HPLC (C18 column, 10%-90% acetonitrile/watergradient).

[0391] Analog 90 (TFA Salt). ¹H NMR (400 MHz, MeOD) δ 7.79 (s, 1H),7.36-7.42 (m, 5H), 7.25-7.32 (m, 2H), 4.14 (d, J=13 Hz, 1H), 3.86 (t,J=11 Hz, 1), 3.62(d,J=11 Hz, 1H) 3.32-3.52(m, 3H),2.81-3.14(m, 13H),2.51(dd, J=12, J=14 Hz, 2H), 2.20 (m, 3H), 1.68-1.87 (m, 3H), 1.50 (m, 3H);ESI (MH⁺) m/z 525.

[0392] Analog 29 (TFA Salt). ¹H NMR (400 MHz, MeOD) δ 7.79 (s, 1H),7.36-7.42 (m, 5H), 7.25-7.32 (m, 2H), 4.14 (d, J=13 Hz, 1H), 3.86 (t,J=11 Hz, 1H), 3.62 (d, J=11 Hz, 1H 3.32-3.52 (m, 5H), 2.81-3.14 (m, 8H),2.51 (m, 3H), 2.20 (m, 2H), 1.68-1.87 (m, 4H), 1.50 (m, 3H), 1.42 (m,6H). ESI (MH⁺) m/z 553.

Example 79

[0393]

[0394] Analog 91 (TFA Salt). ¹H NMR (400 MHz, MeOD) δ 7.79 (s, 1H), 7.40(d, J=8 Hz, 1H), 7.29 (d, J=8 Hz, 1H), 4.25 (q, J=7 Hz, 2H), 3.88 (d,J=12 Hz, 1H), 3.67 (d, J=12 Hz, 1H), 3.57 (m, 1H), 3.10-3.21 (m, 2H),3.02 (t, J=14 Hz, 1H), 2.80-2.93 (m, 3H), 2.53 (dd, J=12, J=14 Hz, 1H),2.19-2.25 (m, 3H), 1.99 (m, 2H), 1.86 (m, 3H), 1.62-1.70 (m. 2H), 1.49(d, J=7 Hz, 3H), 1.24-1.38 (m, 7H); ESI (MH⁺) m/z 507.

[0395] Intermediate 92. SO₃ pyridine complex (42 g, 2.7 mmol) was addedto a DMSO/Et₃N (2.5:1, 0.2 M) solution containing compound 91 (0.34 g,0.65 mmol) at room temperature. After stirring for 2 h, the mixture waspoured into water (25 mL) and extracted with dichloromethane (3×75 mL).The organic layers were washed with brine, dried over Na₂SO₃, andconcentrated to give intermediate 92. Compound 31 was used in the nextstep without purification: ESI (MH⁺) m/z 505.

[0396] Analog 93 (TFA Salt). Sodium triacetoxyborohydride (3 equiv.) wasadded to a dichloromethane solution (0.1 M) containing benzylamine (2equiv.) and the ketone 92 (1 equiv.) at room temperature. After stirringovernight the solvent was removed using evaporation, and the remainingresidue was purified using preparative HPLC (C18 column, 10%-90%acetonitrile/water gradient). ¹H NMR (400 MHz, MeOD,) δ 7.79 (s, 1H),7.44-7.53 (m, 5H), 7.40 (d, J=8 Hz, 1H), 7.29 (d, J=8 Hz, 1H), 4.25 (m,4H), 3.88 (t, J=15 Hz, 1H). 3.75 (d, J=14 Hz, 1H), 3.60 (d, J=14 Hz,1H), 2.75-3.26 (m, 7H), 2.55 (m, 1H), 2.41 (m, 1H), 2.03-2.26 (m, 5H),1.5-1.9 (m, 7H), 1.40-1.50 (m, 4H), 1.31 (m, 3H). ESI (MH⁺) m/z 596.5.

Example 80

[0397] The MCHR modulatory activity of the compounds of the inventioncan be assessed using the in vitro and in vivo assay methods describedabove.

[0398] Exemplary in vitro methods include fluorometric imaging platereader (FLIPR) functional assays (see, e.g., G Protein-Coupled Receptors(1999) pp. 105-108 (T. Haga, G. Bernstein, eds.) CRC Press; Lembo et al.(1999) Nature Cell Biol. 1:267-271; Saito et al. (1999) Nature400:265-269; Wood et al. (2000) Eur. J. Pharmacol. 396:1-8 and Miller etal. (1999) J. Biomol. Screen. 4:249-258) and radioligand binding assays(see, e.g., Receptor Binding Techniques (1999) pp. 37-47 (M. Keen, ed.)Humana Press; Buckley et al. (1989) Mol. Pharmacol. 35:469-476; Miharaet al. (1994) J. Pharmacol. Exp. Ther. 268:1122-1128; Newman et al.(2000) Eur. J. Pharmacol. 397:255-262 and Audinot et al. (2001) Br. J.Pharmacol. 133:371-378).

[0399] Exemplary compounds demonstrated MCHR1 modulatory activity.

[0400] All publications and patent applications cited in thisspecification are herein incorporated by reference as if each individualpublication or patent application were specifically and individuallyindicated to be incorporated by reference. Although the foregoinginvention has been described in some detail by way of illustration andexample for purposes of clarity of understanding, it will be readilyapparent to those of ordinary skill in the art in light of the teachingsof this invention that certain changes and modifications may be madethereto without departing from the spirit or scope of the appendedclaims.

What is claimed is:
 1. A compound having the formula (I):

wherein

represents a single or fused aryl or heteroaryl ring; Q is —N(R)— or—N(R)—(C₁-C₃)alkylene-; R is

L¹ is a bond, (C₁-C₄)alkylene, (C₁-C₄)alkylenoxy and(C₁-C₄)alkylenamino; L² is a bond, (C₁-C₄)alkylene, (C₂-C₄)alkenylene,(C₂-C₄)alkynylene, (C₁-C₄)alkylenoxy (e.g. —OCH₂CH₂—) or(C₁-C₄)alkylenamino (e.g. —NH—CH₂CH₂—); R″ is hydrogen or (C₁-C₈)alkyl;each R¹ is independently selected from the group consisting of halogen,(C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl, fluoro(C₁-C₄)alkyl, —OR⁵,—SR⁵, fluoro(C₁-C₄)alkoxy, aryl, aryl(C₁-C₄)alkyl, —NO₂, —NR⁵R⁶,—C(O)R⁵, —CO₂R⁵, —C(O)NR⁵R⁶, —N(R⁶)C(O)R⁵, —N(R⁶)CO₂R⁵, —N(R⁷)C(O)NR⁵R⁶,—S(O)_(m)NR⁵R⁶, —S(O)_(m)R⁵, —CN and —N(R⁶)S(O)_(m)R⁵; R² and R³ areindependently selected from the group consisting of hydrogen, halogen,(C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl, fluoro(C₁-C₄)alkyl, —OR⁸,—SR⁸, fluoro(C₁-C₄)alkoxy, aryl, aryl(C₁-C₄)alkyl, —NO₂, —NR⁸R⁹, ═O,—C(O)R⁸, —CO₂R⁸, —C(O)NR⁸R⁹, —N(R⁹)C(O)R⁸, —N(R⁹)CO₂R⁸,—N(R¹⁰)C(O)NR⁸R⁹, —S(O)_(m)NR⁸R⁹, —S(O)_(m)R⁸, —CN and —N(R⁹)S(O)_(m)R⁸;R⁴ is selected from the group consisting of hydrogen, —OR¹¹, —C(O)R¹¹,—CO₂R¹¹, —C(O)NR¹¹R¹², —CN, (C₁-C₄)alkyl and aryl; X and Y areindependently selected from the group consisting of (C₁-C₈)alkyl,(C₂-C₈)alkenyl, (C₂-C₈)alkynyl, —CO₂R¹³ and —C(O)NR¹³R¹⁴; optionally, Xand Y may be combined to form a 3-, 4-, 5-, 6- or 7-membered ringcontaining from 0 to 2 heteroatoms independently selected from the groupconsisting of N, O and S; Z is selected from the group consisting of—OR¹⁵, —NR¹⁵R¹⁶, —NR¹⁵R¹⁸, —C(O)R¹⁵, —CO₂R¹⁵, —R¹⁸, —C(O)NR¹⁵R¹⁶,—C(O)NR¹⁵R¹⁸, —SO₂NR¹⁵R¹⁶, —SO₂NR¹⁵R¹⁸, —NR¹⁶SO₂R¹⁵,—N(R¹⁵)N(R¹⁶)SO₂R¹⁷, —C(O)N(R¹⁶)OR¹⁵, hydroxy(C₁-C₈)alkyl,fluoro(C₁-C₄)alkyl, heteroaryl, —C(═NOR¹⁵)NR¹⁶R¹⁷, —C(R¹⁶))═NOR¹⁵,—NR¹⁶(OR¹⁵), —C(O)NR¹⁷C(O)NR¹⁵R¹⁶, —NR¹⁷C(O)NR¹⁶C(O)R¹⁵ and—NR¹⁷C(O)NR¹⁵R¹⁶; R⁵, R⁶, R⁷, R⁸,R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶and R¹⁷ are independently selected from the group consisting ofhydrogen, (C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl,cyclo(C₃-C₆)alkyl, fluoro(C₁-C₄)alkyl, hetero(C₁-C₄)alkyl,cyclohetero(C₃-C₆)alkyl, aryl and aryl(C₁-C₄)alkyl; R¹⁸ is a 5- or6-membered ring containing from 0 to 4 heteroatoms selected from thegroup consisting of N, O and S (e.g. tetrazole); optionally, when two Rgroups selected from the group consisting of R⁵, R⁶, R⁸, R⁹, R¹¹, R¹²,R¹³, R¹⁴, R¹⁵, R¹⁶ and R¹⁷ are attached to the same nitrogen atom, the Rgroups may be combined to form a 3-, 4-, 5-, 6- or 7-membered ringcontaining the nitrogen atom and from 0 to 2 additional heteroatomsselected from the group consisting of N, O and S; the subscript m is 1or 2; and the subscript n is 0, 1 or
 2. 2. The compound of claim 1wherein

represents a benzene ring.
 3. The compound of claim 1 wherein Q is—N(R)—.
 4. The compound of claim 1 wherein R³ is hydrogen or ═O.
 5. Thecompound of claim 1 wherein

represents a benzene ring, R″ is hydrogen and R³ is hydrogen.
 6. Acompound having the formula (II):

or a pharmaceutically acceptable salt, hydrate, solvate or prodrugthereof, wherein L¹ is a bond, (C₁-C₄)alkylene, (C₁-C₄)alkylenoxy or(C₁-C₄)alkylenamino; L² is a bond, (C₁-C₄)alkylene, (C₂-C₄)alkenylene,(C₂-C₄)alkynylene, (C₁-C₄)alkylenoxy or (C₁-C₄)alkylenamino; R″ ishydrogen or (C₁-C₈)alkyl; each R¹ is independently selected from thegroup consisting of halogen, (C₁-C₈)alkyl, (C₂-C₈)alkenyl,(C₂-C₈)alkynyl, fluoro(C₁-C₄)alkyl, —OR⁵, —SR⁵, fluoro(C₁-C₄)alkoxy,aryl, aryl(C₁-C₄)alkyl, —NO₂, —NR⁵R⁶, —C(O)R⁵, —CO₂R⁵, —C(O)NR⁵R⁶,—N(R⁶)C(O)R⁵, —N(R⁶)CO₂R⁵, —N(R⁷)C(O)NR⁵R⁶, —S(O)_(m)NR⁵R⁶, —S(O)_(m)R⁵,—CN and —N(R⁶)S(O)_(m)R⁵; R² is selected from the group consisting ofhydrogen, halogen, (C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl,fluoro(C₁-C₄)alkyl, —OR⁸, —SR⁸, fluoro(C₁-C₄)alkoxy, aryl,aryl(C₁-C₄)alkyl, —NO₂, —NR⁸R⁹, ═O, —C(O)R⁸, —CO₂R⁸, —C(O)NR⁸R⁹,—N(R⁹)C(O)R⁸, —N(R⁹)CO₂R⁸, —N(R¹⁰)C(O)NR⁸R⁹, —S(O)_(m)NR⁸R⁹,—S(O)_(m)R⁸, —CN and —N(R⁹)S(O)_(m)R⁸; R⁴ is selected from the groupconsisting of hydrogen, —OR¹¹, —C(O)R¹¹, —CO₂R¹¹, —C(O)NR¹¹R¹², —CN,(C₁-C₄)alkyl and aryl; X and Y are independently selected from the groupconsisting of (C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl, —CO₂R¹³ and—C(O)NR¹³R¹⁴; optionally, X and Y may be combined to form a 3-, 4-, 5-,6- or 7-membered ring containing from 0 to 2 heteroatoms selected fromthe group consisting of N, O and S; Z is selected from the groupconsisting of —OR¹⁵, —NR¹⁵R¹⁶, —CO₂R¹⁵, —R¹⁸, —C(O)NR¹⁵R¹⁶, —C(O)NR⁵R¹⁸,—SO₂NR¹⁵R¹⁶, —SO₂NR¹⁵R¹⁸, —NR¹⁶SO₂R¹⁵, —N(R¹⁵)N(R¹⁶)SO₂R¹⁷,—C(O)N(R¹⁶)OR¹⁵, fluoro(C₁-C₄)alkyl, heteroaryl, —C(═NOR¹⁵)NR¹⁶R¹⁷,—C(R¹⁶)═NOR¹⁵, —NR¹⁶(OR¹⁵), —C(O)NR¹⁷C(O)NR¹⁵R¹⁶, —NR¹⁷C(O)NR¹⁶C(O)R¹⁵and —NR¹⁷C(O)NR¹⁵R¹⁶; R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵,R¹⁶ and R¹⁷ we independently selected from the group consisting ofhydrogen, (C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl,fluoro(C₁-C₄)alkyl, hetero(C₁-C₄)alkyl, aryl and aryl(C₁-C₄)alkyl; R¹⁸is a 5- or 6-membered ring containing from 1 to 3 heteroatoms selectedfrom the group consisting of N, O and S; optionally, when two R groupsselected from the group consisting of R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹²,R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷ and R¹⁸ are attached to the same nitrogen atom,the R groups may be combined to form a 3-, 4-, 5-, 6- or 7-membered ringcontaining the nitrogen atom and from 0 to 2 additional heteroatomsselected from the group consisting of N, O and S; the subscript m is 1or 2; and the subscript n is 0, 1 or
 2. 7. The compound of claim 6,wherein R⁴ is hydrogen.
 8. The compound of claim 6, wherein R″ ishydrogen.
 9. The compound of claim 8, wherein R² is (C₁-C₄)alkyl oraryl.
 10. The compound of claim 9, wherein R¹ is independently selectedfrom the group consisting of halogen, (C₁-C₄)alkyl, fluoro(C₁-C₄)alkyl,—OR⁵, fluoro(C₁-C₄)alkoxy, —CO₂R⁵, —S(O)_(m)NR⁵R⁶, —S(O)_(m)R⁵ and —CN.11. The compound of claim 10, wherein R¹ is halogen orfluoro(C₁-C₄)alkyl.
 12. The compound of claim 10, wherein n is 0 or 1.13. The compound of claim 12, wherein L¹ is (C₁-C₄)alkylene.
 14. Thecompound of claim 13, having the formula (III):

wherein the subscript p is an integer of from 1 to
 4. 15. The compoundof claim 13, wherein p is 1, 2 or
 3. 16. The compound of claim 15,wherein L² is a bond.
 17. The compound of claim 16, wherein Z is —CO₂R¹⁵or —CO₂NR¹⁵R¹⁶.
 18. The compound of claim 15, wherein X and Y arecombined to form a 3-, 4-, 5-, 6- or 7-membered ring containing from 0to 2 heteroatoms selected from the group consisting of O, N and S. 19.The compound of claim 18, wherein X and Y are combined to form a 5- or6-membered ring containing from 0 to 2 heteroatoms selected from thegroup consisting of O, N and S.
 20. The compound of claim 19, wherein Xand Y are combined to form a 5- or 6-membered ring containing 0heteroatoms, 1 nitrogen atom or 1 oxygen atom.
 21. The compound of claim6, having the formula (IV):

wherein the subscript p is an integer of from 1 to
 4. 22. The compoundof claim 21, wherein p is 1, 2 or
 3. 23. The compound of claim 22,wherein p is
 2. 24. The compound of claim 23, wherein Y is —CO₂H. 25.The compound of claim 23, wherein X and Y are combined to form a 3-, 4-,5-, 6- or 7-membered ring containing from 0 to 2 heteroatoms selectedfrom the group consisting of O, N and S.
 26. The compound of claim 23,wherein X and Y are combined to form a 5- or 6-membered ring containingfrom 0 to 2 heteroatoms selected from the group consisting of O, N andS.
 27. The compound of claim 23, wherein X and Y are combined to form a5- or 6-membered ring containing 0 heteroatoms, 1 nitrogen atom or 1oxygen atom.
 28. The compound of claim 23, wherein X and Y are combinedto form a 5- or 6-membered ring containing 0 heteroatoms, 1 nitrogenatom or 1 oxygen atom and Y is —CO₂H.
 29. The compound of claim 23,wherein R² is methyl.
 30. The compound of claim 23, wherein R¹ is CF₃.31. The compound of claim 30, wherein R¹ is 9-trifluoromethyl.
 32. Thecompound of claim 23, wherein R¹ is CF₃ and R² is methyl.
 33. Thecompound of claim 23, wherein R¹ is CF₃, R² is methyl and Y is —CO₂H.34. The compound of claim 33, wherein said compound is selected from thegroup consisting of the group consisting of:


35. A pharmaceutical composition comprising a pharmaceuticallyacceptable carrier or excipient and a compound of any one of claims1-34.
 36. A method for treating a condition or disorder is selected fromthe group consisting of obesity, an eating disorder, an anxiety disorderand a mood disorder, comprising administering to a subject in needthereof a therapeutically effective amount of a compound of claim 1 or6.
 37. The method of claim 36, wherein said compound condition ordisorder is selected from the group consisting of obesity, anorexianervosa, anxiety, panic disorder and obsessive-compulsive disorder anddepression.
 38. The method of claim 36, wherein said compound isadministered in combination with an anti-obesity agent, anantidepressant or an anxiolytic agent.
 39. The method of claim 36,wherein said compound is administered orally.
 40. The method of claim36, wherein said compound is administered parenterally.
 41. The methodof claim 36, wherein said compound modulates MCHR.
 42. A method formodifying eating behavior, comprising administering to a subject in needthereof a therapeutically effective amount of a compound of claim 1 or6.
 43. The method of claim 42, wherein food intake is decreased.
 44. Themethod of claim 42, wherein food intake is increased.
 45. A method fortreating a condition or disorder mediated by MCHR, comprisingadministering to a subject in need thereof a therapeutically effectiveamount of a compound of claim 1 or
 6. 46. The method of claim 45,wherein said condition or disorder is selected from the group consistingof obesity, an eating disorder, an anxiety disorder and a mood disorder.47. The method of claim 46, wherein said eating disorder is anorexianervosa.
 48. The method of claim 46, wherein said anxiety disorder isselected from the group consisting of anxiety, panic disorder andobsessive-compulsive disorder.
 49. The method of claim 46, wherein saidmood disorder is depression.
 50. A method for modulating MCHR,comprising contacting a cell with a compound of claim 1 or
 6. 51. Themethod of claim 50, wherein said compound is an MCHR antagonist.
 52. Themethod of claim 50, wherein said compound is an MCHR agonist.